Methods for improving kidney function

ABSTRACT

The invention relates to methods for improving renal (kidney) function. Methods for decreasing blood urea nitrogen (BUN), for increasing glomerular filtration rate (GFR), and for decreasing serum creatinine are also provided.

This application is a continuation-in-part application of U.S. patentapplication Ser. No. 10/313,551, filed 6 Dec. 2002, incorporated byreference herein in its entirety.

FIELD OF THE INVENTION

The invention relates to methods for improving renal (kidney) function.Methods for decreasing blood urea nitrogen (BUN), for increasingglomerular filtration rate (GFR), and for decreasing serum creatinineare also provided.

BACKGROUND

The kidneys remove waste product from the body, and regulate and balancebody levels of various fluids and metabolites. Impairment in kidneyfunction can lead to serious health problems, and can be fatal.Decreased or weakened kidney function can be caused by a number ofdifferent factors and conditions, including, although not limited to,diabetes, hypertension, certain drugs or toxins, congenital disease,trauma (including dehydration, shock, etc.), injury, and surgery. Theseand other conditions can ultimately result in, not only weakened orimpaired kidney function, but kidney failure. Subjects with acute orchronic kidney failure may be treated with hemodialysis, peritonealdialysis or kidney transplantation. It is estimated that at least 20million Americans suffer from chronic kidney disease, and another 20million are at risk. (Source: National Kidney Foundation website.)

Improving kidney function in subjects with impaired kidney functionwould contribute greatly to quality of life by reducing the need forserious medical procedures and preventing progression to kidney failure.Therefore, there is a need in the art for methods for improving kidneyfunction. The present invention meets this need by providing methods forimproving kidney function and compounds for use in these methods. Inparticular, the present methods and compounds can be used to improvekidney function in a subject having or at risk for having impairedkidney function. The subject can be a subject having or at risk forhaving acute or chronic kidney failure, or any other impairment of thenormal functioning of the kidney. The present methods and compounds canfurther be used to improve measurable parameters of kidney function,such as serum creatinine, glomerular filtration rate, and blood ureanitrogen (BUN).

SUMMARY OF THE INVENTION

The present invention relates to methods and compounds for improvingkidney function in a subject in need thereof, e.g., a subject having orat risk for having reduced or impaired kidney function. In certainaspects, the improving comprises returning subjects to baseline renalfunction as indicated by serum creatinine levels and other measurableindicators of kidney function.

In one embodiment, the present invention provides a method for improvingkidney function in a subject having or at risk for having impairedkidney function, the method comprising administering to the subject aneffective amount of an agent that inhibits hypoxia inducible factor(HIF) hydroxylase activity.

The agent used in the present methods can be any agent that inhibits HIFhydroxylase activity, including, e.g., a polynucleotide, e.g. antisensesequence; a polypeptide; an antibody or fragment thereof, a smallmolecule, etc. A preferred agent of the present invention is a smallmolecule compound that inhibits HIF hydroxylase activity. In furtherembodiments, the agent is selected from the group consisting of2-oxoglutarate mimetics, iron chelators, and proline analogs. Agents foruse in the present methods include, but are not limited to, agents ofFormulae I, II, III, and IV.

In a preferred embodiment, the agent is a 2-oxoglutarate mimetic. Suchcompounds may inhibit the target 2-oxoglutarate dioxygenase enzymefamily member competitively with respect to 2-oxoglutarate. (Majamaa etal. (1984) Eur J Biochem 138:239-245; and Majamaa et al., supra.) Incertain embodiments, the 2-oxoglutarate mimetic is selected from thegroup consisting of a compound of Formula I and Formula IV. Inparticular embodiments, the 2-oxoglutarate mimetic is apyridine-2-carboxamide including, but not limited to, various compoundsof Formula I. In particular embodiments, the 2-oxoglutarate mimetic is aquinoline-2-carboxamide including, but not limited to, those of FormulaIa. In particular embodiments, the 2-oxoglutarate is anisoquinoline-3-carboxamide including, but not limited to, those ofFormula Ib.

In further embodiments, an agent for use in the present methods isselected from the group consisting of:4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylic acid,N-((1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-acetic acid,3-{[4-(3,3-dibenzyl-ureido)-benzenesulfonyl]-[2-(4-methoxy-phenyl)-ethyl]-amino}-N-hydroxy-propionamide,7-(4-methyl-piperazin-1-ylmethyl)-5-phenylsulfanylmethyl-quinolin-8-ol,4-nitro-quinolin-8-ol, 5-butoxymethyl-quinolin-8-ol,[(3-hydroxy-pyridine-2-carbonyl)-amino]-acetic acid,N-((3-hydroxy-6-isopropoxy-quinoline-2-carbonyl)-amino)-acetic acid,[(3-hydroxy-6-trifluoromethoxy-quinoline-2-carbonyl)-amino]-acetic acid,N-((6-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid,((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid methyl ester,N-((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid, N-((7-butoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid, 6-cyclohexyl-1-hydroxy-4-methyl-1H-pyridin-2-one,[(6-chloro-3-hydroxy-quinoline-2-carbonyl)-amino]-acetic acid,[(3-methoxy-pyridine-2-carbonyl)-amino]-acetic acid,5-methoxy-4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylic acid,[(1,7-dichloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,{[4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid, and[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid.

While, as discussed herein, a preferred agent of the present inventionis a small molecule compound, it is contemplated herein that inhibitingHIF hydroxylase activity can be accomplished by any of the methodsavailable to and known by those of skill in the art, and can involve useof any agent that interacts with, binds to, or modifies HIFα or factorsthat interact with HIFα, including, e.g., enzymes for which HIFα is asubstrate. In certain aspects, the present invention contemplatesproviding a constitutively stable HIFα variant, e.g., stable HIFmuteins, etc, or a polynucleotide encoding such a variant. In furtheraspects, HIFα is HIF1α, HIF2α, or HIF3α. In a preferred aspect,inhibiting HIF hydroxylase activity comprises administering to thesubject an effective amount of an agent that inhibits HIF prolylhydroxylase activity.

Pharmaceutical compositions or medicaments effective for improvingkidney function are also provided herein. In various embodiments, thecompositions comprise an effective amount of an agent that inhibits HIFhydroxylase activity and a carrier. A pharmaceutical compositioneffective for improving kidney function, the composition comprising aneffective amount of an agent that inhibits HIF hydroxylase activity isspecifically contemplated.

In various embodiments, the agent is administered orally, systemically,by injection, and intravenously.

The invention further provides methods for increasing glomerularfiltration rate (GFR) in a subject in need, e.g., a subject having or atrisk for having a decreased GFR, the method comprising administering tothe subject a therapeutically effective amount of an agent that inhibitsHIF hydroxylase activity.

Glomerular filtration rate (GFR) is the best estimate of kidneyfunction. (Source: National Kidney Foundation website.) Therefore, inone embodiment, the subject is a subject having a decreased GFR, i.e., aGFR lower than normal GFR. In one aspect, the subject is a humansubject. In further aspects, the subject is a human subject having or atrisk for having a GFR selected from the group consisting of: below about116 ml/min/1.73 m²; below about 107 ml/min/1.73 m²; below about 99ml/min/1.73 m²; below about 93 ml/min/1.73 m²; below about 85ml/min/1.73 m²; and below about 75 ml/min/1.73 m².

In one aspect, the subject has a GFR below normal GFR, e.g., below about90 ml/min/1.73 m². Therefore, it is contemplated that a subject having aGFR below about 90 ml/min/1.73 m², below about 60 ml/min/1.73 m², belowabout 30 ml/min/1.73 m², or below about 15 ml/min/1.73 m² is a suitablesubject for treatment with the methods or use of medicaments provided bythe present invention.

In certain aspects, the subject is a human subject, and the presentinvention provides methods for increasing GFR in a subject to a GFRgreater than or equal to a GFR selected from the group consisting of:greater than or equal to 90 ml/min/1.73 m²; 60-89 ml/min/1.73 m²; 30-59ml/min/1.73 m²; 15-29 ml/min/1.73 m²; and less than 15 ml/min/1.73 m².

It is further contemplated, in various embodiments, that the methods forincreasing GFR be applied to increase GFR to a level above about 15ml/min/1.73 m². In another aspect, GFR or eGFR is increased to a levelabove about 30 ml/min/1.73 m². In yet another aspect, GFR is increasedto a level above about 60 ml/min/1.73 m². In yet another aspect, GFR isincreased to a level above about 90 ml/min/1.73 m².

A method for decreasing serum creatinine in a subject in need thereof,the method comprising administering to the subject a therapeuticallyeffective amount of an agent that inhibits HIF hydroxylase activity, isalso contemplated. It is contemplated that the subject is a subjecthaving or at risk for having increased serum creatinine or having or atrisk for having impaired kidney function.

In one embodiment, the present invention provides methods for improvingserum creatinine levels in a subject having or at risk for havingincreased creatinine levels, wherein the subject is a human subject.Normal serum creatinine levels are in the range of about 0.8-1.4 mg/dl(70.4-123.2 micromoles per liter) for adult men and about 0.6-1.0 mg/dl(52.8-88 micromoles per liter) for adult women. Therefore, in variousaspects, the subject is a male human subject having a serum creatininelevel of above about 1.4 mg/dl, above about 1.0 mg/dl, or above about0.8 mg/dl. In another aspect, the subject is a female human subjecthaving a serum creatinine level of above about 1.0 mg/dl, above about0.8 mg/dl, or above about 0.6 mg/dl. In various aspects, the decreasingserum creatinine levels can encompass decreasing serum creatinine levelsto below about 1.4, 1.0, 0.8, and 0.6 mg/dl, respectively.

Methods for decreasing blood urea nitrogen (BUN) in a subject having orat risk for having increased, i.e., higher than normal, BUN, the methodcomprising administering to the subject a therapeutically effectiveamount of an agent that inhibits HIF hydroxylase activity, are alsoencompassed herein. In humans, normal BUN levels typically range from7-20 mg/dL. Therefore, in one aspect, subject is a human subject, andthe decreasing BUN comprises decreasing BUN to a value at or below 20mg/dL. In certain aspects, the subject is a human subject, and theincreased BUN is a BUN selected from the group consisting of: above 20mg/dL, above 30 mg/dL, above 40 mg/dL, above 50 mg/dL, above 60 mg/dL,above 70 mg/dL, and above 80 mg/dL.

In one embodiment, the present invention provides a method for reducingcholesterol in a subject having or at risk for having elevatedcholesterol, the method comprising administering to the subject aneffective amount of an agent that inhibits HIF hydroxylase activity. Ina further embodiment, the subject is human, and the reducing cholesterolcomprises reducing blood cholesterol levels to a level selected from thegroup consisting of: below 200 mg/dL, below 180 mg/dL, below 160 mg/dL,and below 150 mg/dL. In another embodiment, the subject is human and theelevated cholesterol is selected from the group consisting of a bloodcholesterol level above 200 mg/dL above 220 mg/dL, and above 240 mg/dL.

In certain embodiments, the subject is a mammalian subject, including,e.g., a cat, a dog, etc. In preferred embodiments, the subject is ahuman subject.

In various embodiments of the present invention, the subject having orat risk for having impaired kidney function is a subject having or atrisk for having any nephropathy or kidney disease. In certainembodiments, the subject has or is at risk for having diabeticnephropathy.

In one embodiment, the subject having or at risk for having impairedkidney function is a subject having or at risk for having acute kidneyfailure. In another embodiment, the subject is a subject having or atrisk for having chronic kidney failure. In other embodiments, thesubject has or is at risk for having a disorder selected from the groupconsisting of diabetes and hypertension.

In further embodiments of the methods described herein, the presentagents are administered in combination with another therapeutic agenthaving a different mode of action, e.g., an ACE inhibitor (ACEI),angiotensin-II receptor blocker (ARB), statin, diuretic, digoxin,camitine, etc.

These and other embodiments of the subject invention will readily occurto those of skill in the art in light of the disclosure herein, and allsuch embodiments are specifically contemplated.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B set forth data showing the methods and compounds of thepresent invention increased glomerular filtration rate in subjects withchronic kidney disease.

FIGS. 2A and 2B set forth data showing the methods and compounds of thepresent invention decreased serum creatinine levels in an animal modelof impaired renal or kidney function.

FIG. 3 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels and serum creatinine levels in an animalmodel of impaired renal function.

FIG. 4 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels and serum creatinine levels in an animalmodel of impaired renal function.

FIG. 5 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels in an animal model ofimpaired renal function.

FIG. 6 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day three following impaired renalfunction.

FIG. 7 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day seven following impaired renalfunction.

FIG. 8 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day ten following impaired kidneyrenal function.

FIG. 9 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day three followingimpaired renal function.

FIG. 10 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day seven followingimpaired renal function.

FIG. 11 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day ten followingimpaired renal function.

FIG. 12 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day three following cisplatin-inducedrenal injury.

FIG. 13 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day seven following cisplatin-inducedkidney injury.

FIG. 14 sets forth data showing the methods and compounds of the presentinvention decreased BUN levels at day fourteen followingcisplatin-induced kidney injury.

FIG. 15 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day three followingcisplatin-induced renal injury.

FIG. 16 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day seven followingcisplatin-induced kidney injury.

FIG. 17 sets forth data showing the methods and compounds of the presentinvention decreased serum creatinine levels at day fourteen followingcisplatin-induced kidney injury.

FIG. 18 shows increased survivability in animals subjected to renalischemic-reperfusion injury that have been pretreated and consequentlytreated with compounds of the invention relative to untreated andsham-operated controls.

FIGS. 19A and 19B show improvement in kidney function followingischemic-reperfusion injury in animals pretreated with a compound of theinvention relative to untreated controls. FIG. 19A shows lower bloodurea nitrogen levels in treated animals relative to untreated controlsat 3 and 7 days after inducing ischemia-reperfusion injury. FIG. 19Bshows lower blood cholesterol levels in treated animals relative tountreated controls at 3, 7, and 14 days after inducingischemia-reperfusion injury.

DESCRIPTION OF THE INVENTION

Before the present compositions and methods are described, it is to beunderstood that the invention is not limited to the particularmethodologies, protocols, cell lines, assays, and reagents described, asthese may vary. It is also to be understood that the terminology usedherein is intended to describe particular embodiments of the presentinvention, and is in no way intended to limit the scope of the presentinvention as set forth in the appended claims.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural references unlesscontext clearly dictates otherwise. Thus, for example, a reference to “afragment” includes a plurality of such fragments; a reference to an“antibody” is a reference to one or more antibodies and to equivalentsthereof known to those skilled in the art, and so forth.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of ordinary skillin the art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methods,devices, and materials are now described. All publications cited hereinare incorporated herein by reference in their entirety for the purposeof describing and disclosing the methodologies, reagents, and toolsreported in the publications that might be used in connection with theinvention. Nothing herein is to be construed as an admission that theinvention is not entitled to antedate such disclosure by virtue of priorinvention.

The practice of the present invention will employ, unless otherwiseindicated, conventional methods of chemistry, biochemistry, molecularbiology, cell biology, genetics, immunology and pharmacology, within theskill of the art. Such techniques are explained fully in the literature.See, e.g., Gennaro, A. R., ed. (1990) Remington's PharmaceuticalSciences, 18^(th) ed., Mack Publishing Co.; Hardman, J. G., Limbird, L.E., and Gilman, A. G., eds. (2001) The Pharmacological Basis ofTherapeutics, 10^(th) ed., McGraw-Hill Co.; Colowick, S. et al., eds.,Methods In Enzymolog, Academic Press, Inc.; Weir, D. M., and Blackwell,C. C., eds. (1986) Handbook of Experimental Immunology, Vols. I-IV,Blackwell Scientific Publications; Maniatis, T. et al., eds. (1989)Molecular Cloning: A Laboratory Manual, 2^(nd) edition, Vols. I-III,Cold Spring Harbor Laboratory Press; Ausubel, F. M. et al., eds. (1999)Short Protocols in Molecular Biology, 4^(th) edition, John Wiley & Sons;Ream et al., eds. (1998) Molecular Biology Techniques: An IntensiveLaboratory Course, Academic Press; Newton, C. R., and Graham, A., eds.(1997) PCR (Introduction to Biotechniques Series), 2^(nd) ed., SpringerVerlag.

The term “HIFα” refers to the alpha subunit of hypoxia inducible factorprotein. HIFα may be any human or other mammalian protein, or fragmentthereof, including, but not limited to, human HIF-1α (Genbank AccessionNo. Q16665), HIF-2α (Genbank Accession No. AAB41495), and HIF-3α(Genbank Accession No. AAD22668); murine HIF-1α (Genbank Accession No.Q61221), HIF-2α (Genbank Accession No. BAA20130 and AAB41496), andHIF-3α (Genbank Accession No. AAC72734); rat HIF-1α (Genbank AccessionNo. CAA70701), HIF-2α (Genbank Accession No. CAB96612), and HIF-3α(Genbank Accession No. CAB96611); and cow HIF-1α (Genbank Accession No.BAA78675). HIFα may also be any non-mammalian protein or fragmentthereof, including Xenopus laevis HIF-1α (Genbank Accession No.CAB96628), Drosophila melanogaster HIF-1α (Genbank Accession No.JC4851), and chicken HIF-1α (Genbank Accession No. BAA34234). HIFα genesequences may also be obtained by routine cloning techniques, forexample, by using all or part of a HIFα gene sequence described above asa probe to recover and determine the sequence of a HIFα gene in anotherspecies.

Fragments of HIFα include the regions defined by human HIF-1α from aminoacid 401 to 603 (Huang et al., supra), amino acid 531 to 575 (Jiang etal. (1997) J Biol Chem 272:19253-19260), amino acid 556 to 575 (Tanimotoet al., supra), amino acid 557 to 571 (Srinivas et al. (1999) BiochemBiophys Res Commun 260:557-561), and amino acid 556 to 575 (Ivan andKaelin (2001) Science 292:464-468). Further, a fragment of HIFα includesany fragment containing at least one occurrence of the motif LXXLAP,e.g., as occurs in the HIF-1α native sequence at L₃₉₇TLLAP andL₅₅₉EMLAP. Additionally, a fragment of HIFα includes any fragmentretaining at least one functional or structural characteristic of HIFα.

The terms “HIF prolyl hydroxylase” and “HIF PH” refer to any enzymecapable of hydroxylating a proline residue in the HIF protein.Preferably, the proline residue hydroxylated by HIF PH includes theproline found within the motif LXXLAP, e.g., as occurs in the humanHIF-1α native sequence at L₃₉₇TLLAP and L₅₅₉EMLAP. HIF PH includesmembers of the Egl-Nine (EGLN) gene family described by Taylor (2001,Gene 275:125-132), and characterized by Aravind and Koonin (2001, GenomeBiol 2:RESEARCH0007), Epstein et al. (2001, Cell 107:43-54), and Bruickand McKnight (2001, Science 294:1337-1340). Examples of HIF PH enzymesinclude human SM-20 (EGLN1) (GenBank Accession No. AAG33965; Dupuy etal. (2000) Genomics 69:348-54), EGLN2 isoform 1 (GenBank Accession No.CAC42510; Taylor, supra), EGLN2 isoform 2 (GenBank Accession No.NP_(—)060025), and EGLN3 (GenBank Accession No. CAC42511; Taylor,supra); mouse EGLN1 (GenBank Accession No. CAC42515), EGLN2 (GenBankAccession No. CAC42511), and EGLN3 (SM-20) (GenBank Accession No.CAC42517); and rat SM-20 (GenBank Accession No. AAA19321). Additionally,HIF PH may include Caenorhabditis elegans EGL-9 (GenBank Accession No.AAD56365) and Drosophila melanogaster CG1114 gene product (GenBankAccession No. AAF52050). HIF PH also includes any fragment retaining atleast one stuctural or function feature of the foregoing full-lengthproteins, including a fragment having hydroxylase activity.

The terms “amino acid sequence” or “polypeptide” as used herein, e.g.,to refer to HIFα and fragments thereof, or HIF PH and fragments thereof,contemplate an oligopeptide, peptide, or protein sequence, or to afragment of any of these, and to naturally occurring or syntheticmolecules. “Fragments” can refer to any portion of a sequence thatretains at least one structural or functional characteristic of theprotein. Immunogenic fragments or antigenic fragments are fragments ofpolypeptides, preferably, fragments of about five to fifteen amino acidsin length, that retain at least one biological or immunologicalactivity. Where “amino acid sequence” is used to refer to thepolypeptide sequence of a naturally occurring protein molecule, “aminoacid sequence” and like terms are not meant to limit the amino acidsequence to the complete native sequence associated with the recitedprotein molecule.

The term “related proteins” as used herein, for example, to refer toproteins related to HIFα prolyl hydroxylase, encompasses other2-oxoglutarate dioxygenase enzymes, especially those family members thatsimilarly require Fe²⁺, 2-oxoglutarate, and oxygen to maintainhydroxylase activity. Such enzymes include, but are not limited to,e.g., procollagen lysyl hydroxylase, procollagen prolyl 4-hydroxylase,and Factor Inhibiting HIF (FIH), an asparaginyl hydroxylase responsiblefor regulating transactivation of HIFα. (GenBank Accession No. AAL27308;Mahon et al. (2001) Genes Dev 15:2675-2686; Lando et al. (2002) Science295:858-861; and Lando et al. (2002) Genes Dev 16:1466-1471. See, also,Elkins et al. (2002) J Biol Chem C200644200.)

The term “agonist” refers to a molecule that increases or prolongs theduration of the effect of a particular molecule, e.g., an enzyme orprotein, or a particular environment, e.g., hypoxia. Agonists mayinclude proteins, nucleic acids, carbohydrates, or any other moleculesthat modulate the effects of the target molecule.

The term “antagonist” refers to a molecule which decreases the extent orduration of the effect of the biological or immunological activity of aparticular molecule. Antagonists may include proteins, nucleic acids,carbohydrates, antibodies, or any other molecules that decrease theeffect of the target molecule.

The term “microarray” refers to any arrangement of nucleic acids, aminoacids, antibodies, etc., on a substrate. The substrate can be anysuitable support, e.g., beads, glass, paper, nitrocellulose, nylon, orany appropriate membrane, etc. A substrate can be any rigid orsemi-rigid support including, but not limited to, membranes, filters,wafers, chips, slides, fibers, beads, including magnetic or nonmagneticbeads, gels, tubing, plates, polymers, microparticles, capillaries, etc.The substrate can provide a surface for coating and/or can have avariety of surface forms, such as wells, pins, trenches, channels, andpores, to which the nucleic acids, amino acids, etc., may be bound.

The term “excipient” as used herein means an inert or inactive substanceused in the production of pharmaceutical products or other tablets,including without limitation any substance used as a binder,disintegrant, coating, compression/encapsulation aid, cream or lotion,lubricant, parenteral, sweetener or flavoring, suspending/gelling agent,or wet granulation agent. Binders include, e.g., carbopol, povidone,xanthan gum, etc.; coatings include, e.g., cellulose acetate phthalate,ethylcellulose, gellan gum, maltodextrin, etc.;compression/encapsulation aids include, e.g., calcium carbonate,dextrose, fructose dc, honey dc, lactose (anhydrate or monohydrate;optionally in combination with aspartame, cellulose, or microcrystallinecellulose), starch dc, sucrose, etc.; disintegrants include, e.g.,croscarmellose sodium, gellan gum, sodium starch glycolate, etc.; creamsand lotions include, e.g., maltodextrin, carrageenans, etc.; lubricantsinclude, e.g., magnesium stearate, stearic acid, sodium stearylfumarate, etc.; materials for chewable tablets include, e.g., dextrose,fructose dc, lactose (monohydrate, optionally in combination withaspartame or cellulose), etc.; parenterals include, e.g., mannitol,povidone, etc.; plasticizers include, e.g., dibutyl sebacate,polyvinylacetate phthalate, etc.; suspending/gelling agents include,e.g., carrageenan, sodium starch glycolate, xanthan gum, etc.;sweeteners include, e.g., aspartame, dextrose, fructose dc, sorbitol,sucrose dc, etc.; and wet granulation agents include, e.g., calciumcarbonate, maltodextrin, microcrystalline cellulose, etc.

The term “sample” is used herein in its broadest sense. Samples may bederived from any source, for example, from bodily fluids, secretions,tissues, cells, or cells in culture including, but not limited to,saliva, blood, urine, serum, plasma, vitreous, synovial fluid, cerebralspinal fluid, amniotic fluid, and organ tissue (e.g., biopsied tissue);from chromosomes, organelles, or other membranes isolated from a cell;from genomic DNA, cDNA, RNA, mRNA, etc.; and from cleared cells ortissues, or blots or imprints from such cells or tissues. Samples may bederived from any source, such as, for example, a human subject, or anon-human mammalian subject, etc. Also contemplated are samples derivedfrom any animal model of disease. A sample can be in solution or can be,for example, fixed or bound to a substrate. A sample can refer to anymaterial suitable for testing for the presence of HIFα or of fragmentsof HIFα or suitable for screening for molecules that bind to HIFα or tofragments thereof. Methods for obtaining such samples are within thelevel of skill in the art.

The term “subject” is used herein in its broadest sense. Subjects mayinclude isolated cells, either prokaryotic or eukaryotic, or tissuesgrown in culture. Preferably, subjects include animals, particularly amammalian species including rat, rabbit, bovine, ovine, porcine, murine,equine, and primate, particularly human.

Invention

The present invention relates to the discovery that administration ofagents that inhibit HIF hydroxylase activity to subjects having or atrisk for having impaired kidney function effectively improved kidneyfunction in these subjects, as demonstrated by improvement in a seriesof measurable parameters of kidney function, including GFR, serumcreatinine levels, BUN, and cholesterol levels.

Measurement of GFR in human subjects is the best overall index of kidneyfunction in health and disease. (Smith, Diseases of the kidney andurinary tract, In: Structure and Function in Health and Disease, NewYork; Oxford Univ. Press, 1951:836-887.) GFR can be determined bymeasuring the urinary clearance of a filtration marker, such as inulin,iothalamate, or iohexol. It is noted that, in a clinical setting, theterm “GFR” often refers to estimated GFR (eGFR), which is a GFR valueestimated by determining creatinine clearance. For the purposes of thepresent invention, the term “GFR” is used herein in reference to thepresent methods and compounds specifically contemplates both GFRobtained through direct measurement and estimated GFR obtained throughany of the various standard formulas used to estimate GFR. Thus, thepresent methods for “increasing GFR” are methods for increasing GFRobtained through direct meaurement and for increasing eGFR, estimatedusing various formulas.

Creatinine clearance (often expressed as ml/min) can be determined bycomparing the level of creatinine in urine with the creatinine level inblood, usually based on assessments of a 24-hour urine sample and ablood sample drawn at the end of the 24-hour period, and is used toestimate GFR. In clinical practice, creatinine clearance is most oftenestimated from the serum creatinine concentration. Creatinine clearanceis related directly to the urine creatinine excretion and inversely toserum creatinine concentration. Various formulas that provide estimatesof creatinine clearance, and therefore estimates of GFR, usingparameters such as serum creatinine concentration, age, sex, and bodysize, have been developed and are standard in the art. (See, e.g.,Cockcroft and Gault (1976) Nephron 16:31-41; Levey et al (1999) Annalsof Internal Medicine 130:462-470; Rule et al (2004) Ann Intern Med141:929-937.)

Normal GFR or eGFR varies according to age, gender, and body weight. Inyoung adults, GFR is approximately 120-130 ml/min/1.73 m². This valuedeclines with age. A persistently reduced GFR is an indication of renalimpairment, such as chronic kidney disease, and often precedes the onsetof kidney failure. The average values of estimated GFR by decade in thegeneral population are as follows: 20-29 years of age, average estimatedGFR of 116 ml/min/1.73 m²; 30-39 years of age, average estimated GFR of107 ml/min/1.73 m²; 40-49 years of age, average estimated GFR of 99ml/min/1.73 m²; 50-59 years of age, average estimated GFR of 93ml/min/1.73 m²; 60-69 years of age, average estimated GFR of 85ml/min/1.73 m²; and 70 years of age and over, average estimated GFR of75 ml/min/1.73 m². Although age-related decline in GFR (or eGFR) hasbeen considered part of normal aging, decreased GFR (or eGFR) in theelderly is an independent predictor of various adverse outcomes, suchas, for example, cardiovascular disease and death, and there is thus anoutstanding need for methods for increasing GFR or eGFR in suchsubjects.

Kidney or renal disorders can lead to reduced or impaired kidneyfunction, which is associated with alterations in various measurableparameters, including, e.g., a change or decline in GFR or eGFR. Variousstages of chronic kidney disease are associated with GFR or eGFR levelsas follows: stage 1 (kidney damage with normal or increased GFR), GFR oreGFR greater than or equal to 90 ml/min/1.73 m²; stage 2 (kidney damagewith mild decrease in GFR), GFR or eGFR of 60-89 ml/min/1.73 m²; stage 3(kidney damage with moderate decrease in GFR), GFR or eGFR of 30-59ml/min/1.73 m²; stage 4 (kidney damage with severe decrease in GFR), GFRor eGFR of 15-29 ml/min/1.73 m²; and stage 5 (kidney failure), GFR oreGFR less than 15 ml/min/1.73 m².

It is contemplated that the present methods can be applied to improvingrenal function, increasing GFR or eGFR in a subject with any clinicallyaccepted standard of measurement indicative of nephropathy or renaldisease, or a subject at risk for developing such a renal disorder. Incertain embodiments, the subject has chronic kidney disease. In variousembodiments, the subject has stage I kidney disease, stage 2 kidneydisease, stage 3 kidney disease, stage 4 kidney disease, or stage 5kidney disease. It is specifically contemplated that the subject hassevere impairment of renal function or late-stage kidney disease.

In one aspect, the subject has a GFR or eGFR below normal GFR or eGFR,e.g., below about 90 ml/min/1.73 m². Therefore, it is contemplated thata subject having a GFR or eGFR below about 90 ml/min/1.73 m², belowabout 60 ml/min/1.73 m², below about 30 ml/min/1.73 m², or below about15 ml/min/1.73 m² is a suitable subject for treatment with the methodsor use of medicaments provided by the present invention.

It is further contemplated, in various embodiments, that the methods forincreasing GFR or eGFR be applied to increase GFR or eGFR to a levelabove about 15 ml/min/1.73 m². In another aspect, GFR or eGFR isincreased to a level above about 30 ml/min/1.73 m². In yet anotheraspect, GFR or eGFR is increased to a level above about 60 ml/min/1.73m². In yet another aspect, GFR or eGFR is increased to a level aboveabout 90 ml/min/1.73 m².

Serum creatinine levels are an established indicator of impaired kidneyfunction and chronic kidney disease. Serum creatinine levels aredetermined by the rate at which serum creatinine is being removed, andcan serve as a measure of kidney function. As kidney function isimpaired, serum creatinine levels increase. Serum creatinine levels canbe expressed as milligrams per deciliter (mg/dl) or as micromoles perliter (μmol/l). For example, a serum creatinine level of 1 mg/dl is thesame as a serum creatinine level of 88 μmol/l. Normal serum creatininelevels are in the range of about 0.8-1.4 mg/dl (70.4-123.2 micromolesper liter) for adult men and about 0.6-1.0 mg/dl (52.8-88 micromoles perliter) for adult women. Therefore, in various aspects, the inventionprovides methods for decreasing, or methods for manufacture of amedicament suitable for use in decreasing, serum creatinine levels in asubject in need thereof. In one aspect, the subject is a male humansubject having a serum creatinine level of above about 1.4 mg/dl, aboveabout 1.0 mg/dl, or above about 0.8 mg/dl. In another aspect, thesubject is a female human subject having a serum creatinine level ofabove about 1.0 mg/dl, above about 0.8 mg/dl, or above about 0.6 mg/dl.In various aspects, the decreasing serum creatinine levels can encompassdecreasing serum creatinine levels to below about 1.4, 1.0, 0.8, and 0.6mg/dl, respectively.

Elevated blood urea nitrogen (BUN) is an indicator of poor orcompromised kidney function. Normal BUN levels for adult humans rangefrom 7-20 mg/dl. Therefore, in certain embodiments, the inventionprovides methods for reducing BUN levels to below 20 mg/dl are providedherein. A greatly elevated BUN (>60 mg/dl) generally indicates amoderate-to-severe degree of renal failure, and methods and compoundsfor decreasing BUN to levels below 60 mg/dl are specificallycontemplated herein.

High cholesterol levels have been linked to kidney disease and kidneyfailure. Increased cholesterol levels can be indicative of loss orimpairment of renal function. As shown in Example 7 and in FIGS. 19A and19B, methods and compounds of the present invention effectively reducedcholesterol levels in an in vivo model of impaired kidney function/renaldisease. Accordingly, the present invention provides in one aspectmethods for reducing elevated cholesterol in a subject having or at riskfor having elevated cholesterol, the methods comprising administering tothe subject an effective amount of an agent that inhibits HIFhydroxylase activity.

In adult human subjects, blood cholesterol levels are preferably below200 mg/dL. Methods for decreasing cholesterol levels to levels of below200 mg/dL, 180 mg/dL, 160 mg/dL, and 150 mg/dL are specifically providedherein. Blood cholesterol levels of about 200-239 mg/dL represent anincreased risk for heart disease and may be associated with kidneydisease, including progressive chronic kidney failure. Levels above 240mg/dL represent a substantial risk for cardiac disease, and may presentan elevated risk of impaired kidney function and chronic kidney disease.Cholesterol levels can be measured by various methods standard in theart (e.g., fasting blood cholesterol, etc.).

While, as discussed herein, a preferred agent of the present inventionis a small molecule compound, it is contemplated herein that inhibitingHIF hydroxylase activity can be accomplished by any of the methodsavailable to and known by those of skill in the art, and can involve useof any agent that interacts with, binds to, or modifies HIFα or factorsthat interact with HIFα, including, e.g., enzymes for which HIFα is asubstrate. In certain aspects, the present invention contemplatesproviding a constitutively stable HIFα variant, e.g., stable HIFmuteins, etc, or a polynucleotide encoding such a variant. In furtheraspects, HIFα is HIF1α, HIF2α, or HIF3α. In a preferred aspect,inhibiting HIF hydroxylase activity comprises administering to thesubject an effective amount.

The agent used in the present methods can be any agent that inhibits HIFhydroxylase activity, including, e.g., a polynucleotide, e.g. antisensesequence; a polypeptide; an antibody or fragment thereof, a smallmolecule, etc. A preferred agent of the present invention is a smallmolecule compound that inhibits HIF hydroxylase activity. In furtherembodiments, the agent is selected from the group consisting of2-oxoglutarate mimetics, iron chelators, and proline analogs. Agents foruse in the present methods include, but are not limited to, agents ofFormulae I, II, III, and IV.

In a preferred embodiment, the agent is a 2-oxoglutarate mimetic. Suchcompounds may inhibit the target 2-oxoglutarate dioxygenase enzymefamily member competitively with respect to 2-oxoglutarate. (Majamaa etal. (1984) Eur J Biochem 138:239-245; and Majamaa et al., supra.) Incertain embodiments, the 2-oxoglutarate mimetic is selected from thegroup consisting of a compound of Formula I and Formula IV. Inparticular embodiments, the 2-oxoglutarate mimetic is apyridine-2-carboxamide including, but not limited to, various compoundsof Formula I. In particular embodiments, the 2-oxoglutarate mimetic is aquinoline-2-carboxamide including, but not limited to, those of FormulaIa. In particular embodiments, the 2-oxoglutarate is anisoquinoline-3-carboxamide including, but not limited to, those ofFormula Ib. In particular embodiments, the 2-oxoglutarate is acinnoline-3-carboxamide including, but not limited to, those of FormulaIc. In particular embodiments, the 2-oxoglutarate is abeta-carboline-3-carboxamide including, but not limited to, variouscompounds of Formula Id.

As stated above, in certain embodiments, compounds used in the methodsof the invention are selected from a compound of the Formula I

-   -   wherein    -   A is 1,2-arylidene, 1,3-arylidene, 1,4-arylidene; or        (C₁-C₄)-alkylene, optionally substituted by one or two halogen,        cyano, nitro, trifluoromethyl, (C₁-C₆)-alkyl,        (C₁-C₆)-hydroxyalkyl, (C₁-C₆)-alkoxy,        —O—[CH₂]_(x)—C_(f)H_((2f+1−g))Hal_(g), (C₁-C₆)-fluoroalkoxy,        (C₁-C₈)-fluoroalkenyloxy, (C₁-C₈)-fluoroalkynyloxy, —OCF₂Cl,        —O—CF₂—CHFCl; (C₁-C₆)-alkylmercapto, (C₁-C₆)-alkylsulfinyl,        (C₁-C₆)-alkylsulfonyl, (C₁-C₆)-alkylcarbonyl,        (C₁-C₆)-alkoxycarbonyl, carbamoyl, N—(C₁-C₄)-alkylcarbamoyl,        N,N-di-(C₁-C₄)-alkylcarbamoyl, (C₁-C₆)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkyl, phenyl, benzyl, phenoxy, benzyloxy, anilino,        N-methylanilino, phenylmercapto, phenylsulfonyl, phenylsulfinyl,        sulfamoyl, N—(C₁-C₄)-alkylsulfamoyl,        N,N-di-(C₁-C₄)-alkylsulfamoyl; or by a substituted        (C₆-C₁₂)-aryloxy, (C₇-C₁₁)-aralkyloxy, (C₆-C₁₂)-aryl,        (C₇-C₁₁)-aralkyl radical, which carries in the aryl moiety one        to five identical or different substituents selected from        halogen, cyano, nitro, trifluoromethyl, (C₁-C₆)-alkyl,        (C₁-C₆)-alkoxy, —O—[CH₂]_(x)-C_(f)H_((2f+1−g))Hal_(g), —OCF₂Cl,        —O—CF₂—CHFCl, (C₁-C₆)-alkylmercapto, (C₁-C₆)-alkylsulfinyl,        (C₁-C₆)-alkylsulfonyl, (C₁-C₆)-alkylcarbonyl,        (C₁-C₆)-alkoxycarbonyl, carbamoyl, N—(C₁-C₄)-alkylcarbamoyl,        N,N-di-(C₁-C₄)-alkylcarbamoyl, (C₁-C₆)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkyl, sulfamoyl, N—(C₁-C₄)-alkylsulfamoyl,        N,N-di-(C₁-C₄)-alkylsulfamoyl; or wherein A is —CR⁵R⁶ and R⁵ and        R⁶ are each independently selected from hydrogen, (C₁-C₆)-alkyl,        (C₃-C₇)-cycloalkyl, aryl, or a substituent of the α-carbon atom        of an α-amino acid, wherein the amino acid is a natural L-amino        acid or its D-isomer;    -   B is —CO₂H, —NH₂, —NHSO₂CF₃, tetrazolyl, imidazolyl,        3-hydroxyisoxazolyl, —CONHCOR′″, —CONHSOR′″, CONHSO₂R′″, where        R′″ is aryl, heteroaryl, (C₃-C₇)-cycloalkyl, or (C₁-C₄)-alkyl,        optionally monosubstituted by (C₆-C₁₂)-aryl, heteroaryl, OH, SH,        (C₁-C₄)-alkyl, (C₁-C₄)-alkoxy, (C₁-C₄)-thioalkyl,        (C₁-C₄)-sulfinyl, (C₁-C₄)-sulfonyl, CF₃, Cl, Br, F, I, NO2,        —COOH, (C₂-C₅)-alkoxycarbonyl, NH₂, mono-(C₁-C₄-alkyl)-amino,        di-(C₁-C₄-alkyl)-amino, or (C₁-C₄)-perfluoroalkyl;    -   or wherein B is a CO₂-G carboxyl radical, where G is a radical        of an alcohol G-OH in which G is selected from (C₁-C₂₀)-alkyl        radical, (C₃-C₈) cycloalkyl radical, (C₂-C₂₀)-alkenyl radical,        (C₃-C₈)-cycloalkenyl radical, retinyl radical, (C₂-C₂₀)-alkynyl        radical, (C₄-C₂₀)-alkenynyl radical, where the alkenyl,        cycloalkenyl, alkynyl, and alkenynyl radicals contain one or        more multiple bonds; (C₆-C₁₆)-carbocyclic aryl radical,        (C₇-C₁₆)-carbocyclic aralkyl radical, heteroaryl radical, or        heteroaralkyl radical, wherein a heteroaryl radical or        heteroaryl moiety of a heteroaralkyl radical contains 5 or 6        ring atoms; and wherein radicals defined for G are substituted        by one or more hydroxyl, halogen, cyano, trifluoromethyl, nitro,        carboxyl, (C₁-C₁₂)-alkyl, (C₃-C₈)-cycloalkyl,        (C₅-C₈)-cycloalkenyl, (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl,        (C₂-C₁₂)-alkenyl, (C₂-C₁₂)-alkynyl, (C₁-C₁₂)-alkoxy,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkyl, (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        —O—[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), —OCF₂Cl, —OCF₂—CHFCl,        (C₁-C₁₂)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₁₂)-alkenylcarbonyl, (C₂-C₁₂)-alkynylcarbonyl,        (C₁-C₁₂)-alkoxycarbonyl,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyl,        (C₆-C₁₂)-aryloxycarbonyl, (C₇-C₁₆)-aralkoxycarbonyl,        (C₃-C₈)-cycloalkoxycarbonyl, (C₂-C₁₂)-alkenyloxycarbonyl,        (C₂-C₁₂)-alkynyloxycarbonyl, acyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyloxy,        (C₆-C₁₂)-aryloxycarbonyloxy, (C₇-C₁₆)aralkyloxycarbonyloxy,        (C₃-C₈)-cycloalkoxycarbonyloxy, (C₂-C₁₂)-alkenyloxycarbonyloxy,        (C₂-C₁₂)-alkynyloxycarbonyloxy, carbamoyl,        N—(C₁-C₁₂)-alkylcarbamoyl, N.N-di(C₁-C₁₂)-alkylcarbamoyl,        N—(C₃-C₈)-cycloalkyl-carbamoyl, N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl,        N-((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)alkyl)-carbamoyl,        N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₆)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        carbamoyloxy, N—(C₁-C₁₂)-alkylcarbamoyloxy,        N.N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N-((C₁-C₁₀)-alkyl)-carbamoyloxy,        N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N-((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        amino, (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₂-C₁₂)-alkenylamino,        (C₂-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C—C₁₁)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino,        (C₁-C₁₂)-alkoxyamino, (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino,        (C₁-C₁₂)-alkylcarbonylamino, (C₃-C₈)-cycloalkylcarbonylamino,        (C₆-C₁₂)arylcarbonylamino, (C₇-C₁₆)-aralkylcarbonylamino,        (C₁-C₁₂)-alkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-arylcarbonyl-N—(C₁-C₁₀)alkylamino,        (C₇-C₁₁)-aralkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₁-C₁₂)-alkylcarbonylamino-(C₁-C₈)-alkyl,        (C₃-C₈)-cycloalkylcarbonylamino-(C₁-C₈)alkyl,        (C₆-C₁₂)-arylcarbonylamino-(C₁-C₈)-alkyl,        (C₇-C₁₂)-aralkylcarbonylamino(C₁-C₈)-alkyl,        amino-(C₁-C₁₀)-alkyl, N—(C₁-C₁₀) alkylamino-(C₁-C₁₀)-alkyl,        N.N-di-(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,        (C₃-C₈)cycloalkylamino-(C₁-C₁₀)-alkyl, (C₁-C₁₂)-alkylmercapto,        (C₁-C₁₂)-alkylsulfinyl, (C₁-C₁₂)-alkylsulfonyl,        (C₆-C₁₆)-arylmercapto, (C₆-C₁₆)-arylsulfinyl,        (C₆-C₁₂)-arylsulfonyl, (C₇-C₁₆)-aralkylmercapto,        (C₇-C₁₆)-aralkylsulfinyl, (C₇-C₁₆)-aralkylsulfonyl, sulfamoyl,        N—(C₁-C₁₀)-alkylsulfamoyl, N.N-di(C₁-C₁₀)-alkylsulfamoyl,        (C₃-C₈)-cycloalkylsulfamoyl, N—(C₆-C₁₂)-alkylsulfamoyl,        N—(C₇-C₁₆)-aralkylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,        (C₁-C₁₀)-alkylsulfonamido,        N-((C₁-C₁₀)-alkyl)-(C₁-C₁₀)-alkylsulfonamido,        (C₇-C₁₆)-aralkylsulfonamido, or        N-((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; wherein radicals        which are aryl or contain an aryl moiety, may be substituted on        the aryl by one to five identical or different hydroxyl,        halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₁-C₁₂)-alkyl, (C₃-C₈)-cycloalkyl, (C₆-C₁₂)-aryl,        (C₇-C₁₆)-aralkyl, (C₁-C₁₂)-alkoxy,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)alkyl, (C₁-C₁₂)-alkoxy-(C₁-C₁₂)alkoxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        (C₁-C₁₂)-alkylcarbonyl, (C₃-C₈)-cycloalkyl-carbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆) aralkylcarbonyl,        (C₁-C₁₂)-alkoxycarbonyl,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyl,        (C₆-C₁₂)-aryloxycarbonyl, (C₇-C₁₆)-aralkoxycarbonyl,        (C₃-C₈)-cycloalkoxycarbonyl, (C₂-C₁₂)-alkenyloxycarbonyl,        (C₂-C₁₂)-alkynyloxycarbonyl, (C₁-C₁₂)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkylcarbonyloxy, (C₆-C₁₂)-arylcarbonyloxy,        (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,        (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyloxy,        (C₆-C₁₂)-aryloxycarbonyloxy, (C₇-C₁₆)-aralkyloxycarbonyloxy,        (C₃-C₈)-cycloalkoxycarbonyloxy, (C₂-C₁₂)-alkenyloxycarbonyloxy,        (C₂-C₁₂)-alkynyloxycarbonyloxy, carbamoyl,        N—(C₁-C₁₂)-alkylcarbamoyl, N.N-di-(C₁-C₁₂)-alkylcarbamoyl,        N—(C₃-C₈)-cycloalkylcarbamoyl, N—(C₆-C₁₂)-arylcarbamoyl,        N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl,        N—((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N—((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        carbamoyloxy, N—(C₁-C₁₂)-alkylcarbamoyloxy,        N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—((C₁-C₁₀)-alkyl)-carbamoyloxy,        N—((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,        amino, (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,        (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C₇-C₁₁)-aralkylamino, N-alkylaralkylamino, N-alkyl-arylamino,        (C₁-C₁₂)-alkoxyamino, (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino,        (C₁-C₁₂)-alkylcarbonylamino, (C₃-C₈)-cycloalkylcarbonylamino,        (C₆-C₁₂)-arylcarbonylamino, (C₇-C₁₆)-alkylcarbonylamino,        (C₁-C₁₂)-alkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-arylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₇-C₁₁)-aralkylcarbonyl-N—(C₁-C₁₀)-alkylamino,        (C₁-C₁₂)-alkylcarbonylamino-(C₁-C₈)-alkyl,        (C₃-C₈)-cycloalkylcarbonylamino-(C₁-C₈)-alkyl,        (C₆-C₁₂)-arylcarbonylamino-(C₁-C₈)-alkyl,        (C₇-C₁₆)-aralkylcarbonylamino-(C₁-C₈)-alkyl,        amino-(C₁-C₁₀)-alkyl, N—(C₁-C₁₀)-alkylamino-(C₁-C₁₀)alkyl,        N.N-di-(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,        (C₃-C₈)-cycloalkylamino-(C₁-C₁₀)-alkyl, (C₁-C₁₂)-alkylmercapto,        (C₁-C₁₂)-alkylsulfinyl, (C₁-C₁₂)-alkylsulfonyl,        (C₆-C₁₂)-arylmercapto, (C₆-C₁₂)-arylsulfinyl,        (C₆-C₁₂)-arylsulfonyl, (C₇-C₁₆)-aralkylmercapto,        (C₇-C₁₆)-aralkylsulfinyl, or (C₇-C₁₆)-aralkylsulfonyl;

X is O or S;

Q is O, S, NR′, or a bond;

where, if Q is a bond, R⁴ is halogen, nitrile, or trifluoromethyl;

or where, if Q is O, S, or NR′, R⁴ is hydrogen, (C₁-C₁₀)-alkyl radical,(C₂-C₁₀)-alkenyl radical, (C₂-C₁₀)-alkynyl radical, wherein alkenyl oralkynyl radical contains one or two C—C multiple bonds; unsubstitutedfluoroalkyl radical of the formula —[CH₂]_(x)-C_(f)H_((2f+1−g))—F_(g),(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl radical,(C₁-C₆)-alkoxy-(C₁-C₄)-alkoxy-(C₁-C₄)-alkyl radical, aryl radical,heteroaryl radical, (C₇-C₁₁)-aralkyl radical, or a radical of theformula Z

—[CH₂]_(v)—[O]_(w)—[CH₂]_(t)—E   (Z)

where

E is a heteroaryl radical, a (C₃-C₈)-cycloalkyl radical, or a phenylradical of the formula F

v is 0-6,

w is 0 or 1,

t is 0-3, and

-   -   R⁷, R⁸, R⁹, R¹⁰, and R¹¹ are identical or different and are        hydrogen, halogen, cyano, nitro, trifluoromethyl, (C₁-C₆)-alkyl,        (C₃-C₈)-cycloalkyl, (C₁-C₆)-alkoxy,        —O—[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), —OCF₂—Cl, —O—CF₂—CHFCl,        (C₁-C₆)-alkylmercapto, (C₁-C₆)-hydroxyalkyl,        (C₁-C₆)-alkoxy-(C₁-C₆)-alkoxy, (C₁-C₆)-alkoxy-(C₁-C₆)-alkyl,        (C₁-C₆)-alkylsulfinyl, (C₁-C₆)-alkylsulfonyl,        (C₁-C₆)-alkylcarbonyl, (C₁-C₈)-alkoxycarbonyl, carbamoyl,        N—(C₁-C₈)-alkylcarbamoyl, N,N-di-(C₁-C₈)-alkylcarbamoyl, or        (C₇-C₁₁)-aralkylcarbamoyl, optionally substituted by fluorine,        chlorine, bromine, trifluoromethyl, (C₁-C₆)-alkoxy,        N—(C₃-C₈)-cycloalkylcarbamoyl,        N—(C₃-C₈)-cycloalkyl-(C₁-C₄)-alkylcarbamoyl,        (C₁-C₆)-alkylcarbonyloxy, phenyl, benzyl, phenoxy, benzyloxy,        NR^(Y)R^(Z) wherein R^(y) and R^(z) are independently selected        from hydrogen, (C₁-C₁₂)-alkyl, (C₁-C₈)-alkoxy-(C₁-C₈)-alkyl,        (C₇-C₁₂)-aralkoxy-(C₁-C₈)-alkyl, (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkyl,        (C₃-C₁₀)-cycloalkyl, (C₃-C₁₂)-alkenyl, (C₃-C₁₂)-alkynyl,        (C₆-C₁₂)-aryl, (C₇-C₁₁)-aralkyl, (C₁-C₁₂)-alkoxy,        (C₇-C₁₂)aralkoxy, (C₁-C₁₂)-alkylcarbonyl,        (C₃-C₈)-cycloalkylcarbonyl, (C₆-C₁₂)arylcarbonyl,        (C₇-C₁₆)-aralkylcarbonyl; or further wherein R^(y) and R^(z)        together are —[CH2]_(h), in which a CH₂ group can be replaced by        O, S, N—(C₁-C₄)-alkylcarbonylimino, or        N—(C₁-C₄)-alkoxycarbonylimino; phenylmercapto, phenylsulfonyl,        phenylsulfinyl, sulfamoyl, N—(C₁-C₈)-alkylsulfamoyl, or        N,N-di-(C₁-C₈)-alkylsulfamoyl; or alternatively R⁷ and R⁸, R⁸        and R⁹, R⁹ and R¹⁰, or R¹⁰ and R¹¹, together are a chain        selected from —[CH₂]_(n)— or —CH═CH—CH═CH—, where a CH₂ group of        the chain is optionally replaced by O, S, SO, SO₂, or NR^(Y);        and n is 3, 4, or 5; and if E is a heteroaryl radical, said        radical can carry 1-3 substituents selected from those defined        for R⁷—R¹¹, or if E is a cycloalkyl radical, the radical can        carry one substituent selected from those defined for R⁷—R¹¹;

or where, if Q is NR′, R⁴ is alternatively R″, where R′ and R″ areidentical or different and are hydrogen, (C₆-C₁₂)-aryl,(C₇-C₁₁)-aralkyl, (C₁-C₈)-alkyl, (C₁-C₈)-alkoxy-(C₁-C₈)-alkyl,(C₇-C₁₂)-aralkoxy-(C₁-C₈)-alkyl, (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkyl,(C₁-C₁₀)-alkylcarbonyl, optionally substituted (C₇-C₁₆)-aralkylcarbonyl,or optionally substituted C₆-C₁₂)-arylcarbonyl; or R′ and R″ togetherare —[CH₂]_(h), in which a CH₂ group can be replaced by O, S,N-acylimino, or N—(C₁-C₁₀)-alkoxycarbonylimino, and h is 3 to 7.

Y is N or CR³;

-   -   R¹, R² and R³ are identical or different and are hydrogen,        hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₁-C₂₀)-alkyl, (C₃-C₈)-cycloalkyl,        (C₃-C₈)cycloalkyl-(C₁-C₁₂)-alkyl, (C₃-C₈)-cycloalkoxy,        (C₃-C₈)-Cycloalkyl-(C₁-C₁₂)-alkoxy,        (C₃-C₈)-cycloalkoxy-(C₁-C₁₂)-alkyl,        (C₃-C₈)-cycloalkyloxy-(C₁-C₁₂)-alkoxy,        (C₃-C₈)-cycloalkyl-(C₁-C₈)-alkyl-(C₁-C₆)-alkoxy,        (C₃-C₈)-cycloalkyl-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,        (C₃-C₈)-cycloalkyloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,        (C₃-C₈)-cycloalkoxy-(C₁-C₈)-alkoxy-(C₁-C₈)-alkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₇-C₁₆)-aralkenyl,        (C₇-C₁₆)-aralkynyl, (C₂-C₂₀)-alkenyl, (C₂-C₂₀)-alkynyl,        (C₁-C₂₀)-alkoxy, (C₂-C₂₀)-alkenyloxy, (C₂-C₂₀)-alkynyloxy,        retinyloxy, (C₁-C₂₀)-alkoxy-(C₁-C₁₂)-alkyl,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxy,        (C₁-C₁₂)-alkoxy-(C₁-C₈)-alkoxy-(C₁-C₈)-alkyl, (C₆-C₁₂)-aryloxy,        (C₇-C₁₆)-aralkyloxy, (C₆-C₁₂)-aryloxy-(C₁-C₆)-alkoxy,        (C₇-C₁₆)-aralkoxy-(C₁-C₆)-alkoxy, (C₁-C₁₆)-hydroxyalkyl,        (C₆-C₁₆)-aryloxy-(C₁-C₈)-alkyl, (C₇-C₁₆)-aralkoxy-(C₁-C₈)-alkyl,        (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,        (C₇-C₁₂)-aralkyloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,        (C₂-C₂₀)-alkenyloxy-(C₁-C₆)-alkyl,        (C₂-C₂₀)-alkynyloxy-(C₁-C₆)-alkyl, retinyloxy-(C₁-C₆)-alkyl,        —O—[CH₂]_(x)CfH_((2f+1−g))F_(g), —OCF₂Cl, —OCF₂—CHFCl,        (C₁-C₂₀)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₂₀)-alkenylcarbonyl, (C₂-C₂₀)-alkynylcarbonyl,        (C₁-C₂₀)-alkoxycarbonyl,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyl,        (C₆-C₁₂)-aryloxycarbonyl, (C₇-C₁₆)-aralkoxycarbonyl,        (C₃-C₈)-cycloalkoxycarbonyl, (C₂-C₂₀)-alkenyloxycarbonyl,        retinyloxycarbonyl, (C₂-C₂₀)-alkynyloxycarbonyl,        (C₆-C₁₂)-aryloxy-(C₁-C₆)-alkoxycarbonyl,        (C₇-C₁₆)-aralkoxy-(C₁-C₆)-alkoxycarbonyl,        (C₃-C₈)-cycloalkyl-(C₁-C₆)-alkoxycarbonyl,        (C₃-C₈)-cycloalkoxy-(C₁-C₆)-alkoxycarbonyl,        (C₁-C₁₂)-alkylcarbonyloxy, (C₃-C₈)-cycloalkylcarbonyloxy,        (C₆-C₁₂)-arylcarbonyloxy, (C₇-C₁₆)-aralkylcarbonyloxy,        cinnamoyloxy, (C₂-C₁₂)-alkenylcarbonyloxy,        (C₂-C₁₂)-alkynylcarbonyloxy, (C₁-C₁₂)-alkoxycarbonyloxy,        (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyloxy,        (C₆-C₁₂)-aryloxycarbonyloxy, (C₇-C₁₆)-aralkyloxycarbonyloxy,        (C₃-C₈)-cycloalkoxycarbonyloxy, (C₂-C₁₂)-alkenyloxycarbonyloxy,        (C₂-C₁₂)-alkynyloxycarbonyloxy, carbamoyl,        N—(C₁-C₁₂)-alkylcarbamoyl, N,N-di-(C₁-C₁₂)-alkylcarbamoyl,        N—(C₃-C₈)-cycloalkylcarbamoyl,        N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,        N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)-carbamoyl,        N—(C₁-C₆)-alkyl-N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)-carbamoyl,        N-(+)-dehydroabietylcarbamoyl,        N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,        N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl,        N—((C₁-C₁₈)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N-((C₆-C₁₆)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N—((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl;        CON(CH₂)_(h), in which a CH₂ group can be replaced by O, S,        N—(C₁-C₈)-alkylimino, N—(C₃-C₈)-cycloalkylimino,        N—(C₃-C₈)-cycloalkyl-(C₁-C₄)-alkylimino, N—(C₆-C₁₂)-arylimino,        N—(C₇-C₁₆)-aralkylimino, N—(C₁-C₄)-alkoxy-(C₁-C₆)-alkylimino,        and h is from 3 to 7; a carbamoyl radical of the formula R

in which

-   -   R^(x) and R^(v) are each independently selected from hydrogen,        (C₁-C₆)-alkyl, (C₃-C₇)-cycloalkyl, aryl, or the substituent of        an α-carbon of an α-amino acid, to which the L- and D-amino        acids belong,

s is 1-5,

-   -   T is OH, or NR*R**, and R*, R** and R*** are identical or        different and are selected from hydrogen, (C₆-C₁₂)-aryl,        (C₇-C₁₁)-aralkyl, (C₁-C₈)-alkyl, (C₃-C₈)-cycloalkyl,        (+)-dehydroabietyl, (C₁-C₈)-alkoxy-(C₁-C₈)-alkyl,        (C₇-C₁₂)-aralkoxy-(C₁-C₈)-alkyl, (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkyl,        (C₁-C₁₀)-alkanoyl, optionally substituted (C₇-C₁₆)-aralkanoyl,        optionally substituted (C₆-C₁₂)-aroyl; or R* and R** together        are —[CH₂]_(h), in which a CH₂ group can be replaced by O, S,        SO, SO₂, N-acylamino, N—(C₁-C₁₀)-alkoxycarbonylimino,        N—(C₁-C₈)-alkylimino, N—(C₃-C₈)-cycloalkylimino,        N—(C₃-C₈)-cycloalkyl-(C₁-C₄)-alkylimino, N—(C₆-C₁₂)-arylimino,        N—(C₇-C₁₆)-aralkylimino, N—(C₁-C₄)-alkoxy-(C₁-C₆)-alkylimino,        and h is from 3 to 7;

-   carbamoyloxy, N—(C₁-C₁₂)-alkylcarbamoyloxy,    N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy, N—(C₃-C₈)-cycloalkylcarbamoyloxy,    N—(C₆-C₁₂)-arylcarbamoyloxy, N—(C₇-C₁₆)-aralkylcarbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,    N—((C₁-C₁₀)-alkyl)-carbamoyloxy,    N—((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,    N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)-carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxyamino,    (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,    (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,    (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,    N—(C₇-C₁₁)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino,    (C₁-C₁₂)-alkoxyamino, (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino,    (C₁-C₁₂)-alkanoylamino, (C₃-C₈)-cycloalkanoylamino,    (C₆-C₁₂)-aroylamino, (C₇-C₁₆)-aralkanoylamino,    (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,    (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₁-C₁₂)-alkanoylamino-(C₁-C₈)-alkyl,    (C₃-C₈)-cycloalkanoylamino-(C₁-C₈)-alkyl,    (C₆-C₁₂)-aroylamino-(C₁-C₈)-alkyl,    (C₇-C₁₆)-aralkanoylamino-(C₁-C₈)-alkyl, amino-(C₁-C₁₀)-alkyl,    N—(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,    N,N-di(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,    (C₃-C₈)-cycloalkylamino(C₁-C₁₀)-alkyl, (C₁-C₂₀)-alkylmercapto,    (C₁-C₂₀)-alkylsulfinyl, (C₁-C₂₀)-alkylsulfonyl,    (C₆-C₁₂)-arylmercapto, (C₆-C₁₂)-arylsulfinyl, (C₆-C₁₂)-arylsulfonyl,    (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl,    (C₇-C₁₂)-aralkylsulfonyl, (C₁-C₁₂)-alkylmercapto-(C₁-C₆)-alkyl,    (C₁-C₁₂)-alkylsulfinyl-(C₁-C₆)-alkyl,    (C₁-C₁₂)-alkylsulfonyl-(C₁-C₆)-alkyl,    (C₆-C₁₂)-arylmercapto-(C₁-C₆)-alkyl,    (C₆-C₁₂)-arylsulfinyl-(C₁-C₆)-alkyl,    (C₆-C₁₂)-arylsulfonyl-(C₁-C₆)-alkyl,    (C₇-C₁₆)-aralkylmercapto-(C₁-C₆)-alkyl,    (C₇-C₁₆)-aralkylsulfinyl-(C₁-C₆)-alkyl,    (C₇-C₁₆)-aralkylsulfonyl-(C₁-C₆)-alkyl, sulfamoyl,    N—(C₁-C₁₀)-alkylsulfamoyl, N,N-di-(C₁-C₁₀)-alkylsulfamoyl,    (C₃-C₈)-cycloalkylsulfamoyl, N—(C₆-C₁₂)-arylsulfamoyl,    N—(C₇-C₁₆)-aralkylsulfamoyl,    N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,    N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,    (C₁-C₁₀)-alkylsulfonamido,    N-((C₁-C₁₀)-alkyl)-(C₁-C₁₀)-alkylsulfonamido,    (C₇-C₁₆)-aralkylsulfonamido, and    N—((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; where an aryl radical    may be substituted by 1 to 5 substituents selected from hydroxyl,    halogen, cyano, trifluoromethyl, nitro, carboxyl, (C₂-C₁₆)-alkyl,    (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkyl-(C₁-C₁₂)-alkyl,    (C₃-C₈)-cycloalkoxy, (C₃-C₈)-cycloalkyl-(C₁-C₁₂)-alkoxy,    (C₃-C₈)-cycloalkyloxy-(C₁-C₁₂)-alkyl,    (C₃-C₈)-Cycloalkyloxy-(C₁-C₁₂)-alkoxy,    (C₃-C₈)-cycloalkyl-(C₁-C₈)-alkyl-(C₁-C₆)-alkoxy,    (C₃-C₈)-cycloalkyl(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,    (C₃-C₈)-cycloalkyloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,    (C₃-C₈)-cycloalkoxy-(C₁-C₈)-alkoxy-(C₁-C₈)-alkoxy, (C₆-C₁₂)-aryl,    (C₇-C₁₆)-aralkyl, (C₂-C₁₆)-alkenyl, (C₂-C₁₂)-alkynyl,    (C₁-C₁₆)-alkoxy, (C₁-C₁₆)-alkenyloxy,    (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkyl, (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxy,    (C₁-C₁₂)-alkoxy(C₁-C₈)-alkoxy-(C₁-C₈)-alkyl, (C₆-C₁₂)-aryloxy,    (C₇-C₁₆)-aralkyloxy, (C₆-C₁₂)-aryloxy-(C₁-C₆)-alkoxy,    (C₇-C₁₆)-aralkoxy-(C₁-C₆)-alkoxy, (C₁-C₈)-hydroxyalkyl,    (C₆-C₁₆)-aryloxy-(C₁-C₈)-alkyl, (C₇-C₁₆)-aralkoxy-(C₁-C₈)-alkyl,    (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,    (C₇-C₁₂)-aralkyloxy-(C₁-C₈)-alkoxy-(C₁-C₆)-alkyl,    —O—[CH₂]_(x)C_(f)H_((2f+1−g))F_(g), —OCF₂Cl, —OCF₂—CHFCl,    (C₁-C₁₂)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,    (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl,    (C₁-C₁₂)-alkoxycarbonyl, (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyl,    (C₆-C₁₂)-aryloxycarbonyl, (C₇-C₁₆)-aralkoxycarbonyl,    (C₃-C₈)-cycloalkoxycarbonyl, (C₂-C₁₂)-alkenyloxycarbonyl,    (C₂-C₁₂)-alkynyloxycarbonyl,    (C₆-C₁₂)-aryloxy-(C₁-C₆)-alkoxycarbonyl,    (C₇-C₁₆)-aralkoxy-(C₁-C₆)-alkoxycarbonyl,    (C₃-C₈)-cycloalkyl-(C₁-C₆)-alkoxycarbonyl,    (C₃-C₈)-cycloalkoxy-(C₁-C₆)-alkoxycarbonyl,    (C₁-C₁₂)-alkylcarbonyloxy, (C₃-C₈)-cycloalkylcarbonyloxy,    (C₆-C₁₂)-arylcarbonyloxy, (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,    (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,    (C₁-C₁₂)-alkoxycarbonyloxy,    (C₁-C₁₂)-alkoxy-(C₁-C₁₂)-alkoxycarbonyloxy,    (C₆-C₁₂)-aryloxycarbonyloxy, (C₇-C₁₆)-aralkyloxycarbonyloxy,    (C₃-C₈)-cycloalkoxycarbonyloxy, (C₂-C₁₂)-alkenyloxycarbonyloxy,    (C₂-C₁₂)-alkynyloxycarbonyloxy, carbamoyl,    N—(C₁-C₁₂)-alkylcarbamoyl, N,N-di(C₁-C₁₂)-alkylcarbamoyl,    N—(C₃-C₈)-cycloalkylcarbamoyl, N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,    N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,    N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)carbamoyl,    N—(C₁-C₆)-alkyl-N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)carbamoyl,    N-(+)-dehydroabietylcarbamoyl,    N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,    N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,    N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,    N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl,    N—((C₁-C₁₆)-alkoxy-(C₁-C₁₀)-alkyl)carbamoyl,    N—((C₆-C₁₆)-aryloxy-(C₁-C₁₀)-alkyl)carbamoyl,    N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)carbamoyl,    N—(C₁-C₁₀)-alkyl-N-((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)carbamoyl,    N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)carbamoyl,    N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,    CON(CH₂)_(h), in which a CH₂ group can be replaced by, O, S,    N—(C₁-C₈)-alkylimino, N—(C₃-C₈)-cycloalkylimino,    N—(C₃-C₈)-cycloalkyl-(C₁-C₄)-alkylimino, N—(C₆-C₁₂)-arylimino,    N—(C₇-C₁₆)-aralkylimino, N—(C₁-C₄)-alkoxy-(C₁-C₆)-alkylimino, and h    is from 3 to 7; carbamoyloxy, N—(C₁-C₁₂)-alkylcarbamoyloxy,    N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy, N—(C₃-C₈)-cycloalkylcarbamoyloxy,    N—(C₆-C₁₆)-arylcarbamoyloxy, N—(C₇-C₁₆)-aralkylcarbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,    N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,    N—((C₁-C₁₀)-alkyl)carbamoyloxy,    N—((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)carbamoyloxy,    N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N-((C₁-C₁₀)-alkoxy-(C₁-C₁₀)-alkyl)carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)carbamoyloxy,    N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)carbamoyloxy,    amino, (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,    (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,    (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino, N-C₇-C₁₁)-aralkylamino,    N-alkyl-aralkylamino, N-alkyl-arylamino, (C₁-C₁₂)-alkoxyamino,    (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino, (C₁-C₁₂)-alkanoylamino,    (C₃-C₈)-cycloalkanoylamino, (C₆-C₁₂)-aroylamino,    (C₇-C₁₆)-aralkanoylamino, (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,    (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino,    (C₁-C₁₂)-alkanoylamino-(C₁-C₈)-alkyl,    (C₃-C₈)-cycloalkanoylamino-(C₁-C₈)-alkyl,    (C₆-C₁₂)-aroylamino-(C₁-C₈)-alkyl,    (C₇-C₁₆)-aralkanoylamino-(C₁-C₈)-alkyl, amino-(C₁-C₁₀)-alkyl,    N—(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,    N,N-di-(C₁-C₁₀)-alkylamino-(C₁-C₁₀)-alkyl,    (C₃-C₈)-cycloalkylamino-(C₁-C₁₀)-alkyl, (C₁-C₁₂)-alkylmercapto,    (C₁-C₁₂)-alkylsulfinyl, (C₁-C₁₂)-alkylsulfonyl,    (C₆-C₁₆)-arylmercapto, (C₆-C₁₆)-arylsulfinyl, (C₆-C₁₆)-arylsulfonyl,    (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl, or    (C₇-C₁₆)-aralkylsulfonyl;    -   or wherein R¹ and R², or R² and R³ form a chain [CH₂]_(o), which        is saturated or unsaturated by a C═C double bond, in which 1 or        2 CH₂ groups are optionally replaced by O, S, SO, SO₂, or NR′,        and R′ is hydrogen, (C₆-C₁₂)-aryl, (C₁-C₈)-alkyl,        (C₁-C₈)-alkoxy-(C₁-C₈)-alkyl, (C₇-C₁₂)-aralkoxy-(C₁-C₈)-alkyl,        (C₆-C₁₂)-aryloxy-(C₁-C₈)-alkyl, (C₁-C₁₀)-alkanoyl, optionally        substituted (C₇-C₁₆)-aralkanoyl, or optionally substituted        (C₆-C₁₂)-aroyl; and o is 3, 4 or 5;    -   or wherein the radicals R¹ and R², or R² and R³, together with        the pyridine or pyridazine carrying them, form a        5,6,7,8-tetrahydroisoquinoline ring, a        5,6,7,8-tetrahydroquinoline ring, or a        5,6,7,8-tetrahydrocinnoline ring;    -   or wherein R¹ and R², or R² and R³ form a carbocyclic or        heterocyclic 5- or 6-membered aromatic ring;    -   or where R¹ and R², or R² and R³, together with the pyridine or        pyridazine carrying them, form an optionally substituted        heterocyclic ring systems selected from thienopyridines,        furanopyridines, pyridopyridines, pyrimidinopyridines,        imidazopyridines, thiazolopyridines, oxazolopyridines,        quinoline, isoquinoline, and cinnoline; where quinoline,        isoquinoline or cinnoline preferably satisfy the formulae Ia, Ib        and Ic:

-   -   and the substituents R¹² to R²³ in each case independently of        each other have the meaning of R¹, R² and R³;    -   or wherein the radicals R¹ and R², together with the pyridine        carrying them, form a compound of Formula Id:

where

-   -   V is S, O, or NR^(k), and R^(k) is selected from hydrogen,        (C₁-C₆)-alkyl, aryl, or benzyl; where an aryl radical may be        optionally substituted by 1 to 5 substituents as defined above;        and

R²⁴, R²⁵, R²⁶, and R²⁷ in each case independently of each other have themeaning of R¹, R² and R³;

f is 1 to 8;

g is 0 or 1 to (2f+1);

x is 0 to 3; and

h is 3 to 7;

including the physiologically active salts and prodrugs derivedtherefrom.

Exemplary compounds according to Formula I are described in EuropeanPatent Nos. EP0650960 and EP0650961. All compounds listed in EP0650960and EP0650961, in particular, those listed in the compound claims andthe final products of the working examples, are hereby incorporated intothe present application by reference herein. Additionally, exemplarycompounds according to Formula I are described in U.S. Pat. No.5,658,933. All compounds listed in U.S. Pat. No. 5,658,933, inparticular, those listed in the compound claims and the final productsof the working examples, are hereby incorporated into the presentapplication by reference herein.

Additional compounds according to Formula I are substituted heterocycliccarboxyamides described in U.S. Pat. No. 5,620,995;3-hydroxypyridine-2-carboxamidoesters described in U.S. Pat. No.6,020,350; sulfonamidocarbonylpyridine-2-carboxamides described in U.S.Pat. No. 5,607,954; and sulfonamidocarbonyl-pyridine-2-carboxamides andsulfonamidocarbonyl-pyridine-2-carboxamide esters described in U.S. Pat.Nos. 5,610,172 and 5,620,996. All compounds listed in these patents, inparticular, those compounds listed in the compound claims and the finalproducts of the working examples, are hereby incorporated into thepresent application by reference herein.

Exemplary compounds according to Formula Ia are described in U.S. Pat.Nos. 5,719,164 and 5,726,305. All compounds listed in the foregoingpatents, in particular, those listed in the compound claims and thefinal products of the working examples, are hereby incorporated into thepresent application by reference herein. Exemplary compounds accordingto Formula Ib are described in U.S. Pat. No. 6,093,730. All compoundslisted in U.S. Pat. No. 6,093,730, in particular, those listed in thecompound claims and the final products of the working examples, arehereby incorporated into the present application by reference herein

Additionally, compounds related to Formula I that can also be used inthe methods of the invention include, but are not limited to,6-cyclohexyl-1-hydroxy-4-methyl-1H-pyridin-2-one (Compound N),7-(4-methyl-piperazin-1-ylmethyl)-5-phenylsulfanylmethyl-quinolin-8-ol(Compound D), 4-nitro-quinolin-8-ol (Compound E), and5-butoxymethyl-quinolin-8-ol (Compound F). Further, the inventionprovides additional exemplary compounds wherein, e.g., position A and Btogether may be, e.g., hexanoic acid, cyanomethyl, 2-aminoethyl, benzoicacid, 1H-benzoimidazol-2-ylmethyl, etc.

In certain embodiments, compounds used in the methods of the inventionare pyridine-2-carboxamides. In one embodiment, the compound is selectedfrom a compound of the Formula I wherein

-   -   A is —CR⁵R⁶—, and R⁵ and R⁶ are each independently selected from        the group consisting of hydrogen, (C₁-C₆)-alkyl,        (C₃-C₇)-cycloalkyl, aryl, or a substituent of the α-carbon atom        of an α-amino acid, wherein the amino acid is a natural L-amino        acid or its D-isomer;    -   B is —CO₂H or a CO₂-G carboxyl radical, where G is a radical of        an alcohol G-OH in which G is selected from the group consisting        of (C₁-C₂₀)-alkyl radical, (C₃-C₈) cycloalkyl radical,        (C₂-C₂₀)-alkenyl radical, (C₃-C₈)-cycloalkenyl radical, retinyl        radical, (C₂-C₂₀)-alkynyl radical, (C₄-C₂₀)-alkenynyl radical;

X is O;

Q is O;

-   -   R⁴ is selected from the group consisting of hydrogen,        (C₁-C₁₀)-alkyl, (C₂-C₁₀)-alkenyl, (C₂-C₁₀)-alkynyl, wherein        alkenyl or alkynyl contains one or two C—C multiple bonds;        unsubstituted fluoroalkyl radical of the formula        —[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), aryl, heteroaryl, and        (C₇-C₁₁)-aralkyl;

Y is CR³;

-   -   R¹, R² and R³ are identical or different and are selected from        the group consisting of hydrogen, hydroxyl, halogen, cyano,        trifluoromethyl, nitro, carboxyl; (C₁-C₂₀)-alkyl,        (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy, (C₆-C₁₂)-aryl,        (C₇-C₁₆)-aralkyl, (C₇-C₁₆)-aralkenyl, (C₇-C₁₆)-aralkynyl,        (C₂-C₂₀)-alkenyl, (C₂-C₂₀)-alkynyl, (C₁-C₂₀)-alkoxy,        (C₂-C₂₀)-alkenyloxy, (C₂-C₂₀)-alkynyloxy, retinyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₁₆)-hydroxyalkyl,        —O—[CH₂]_(x)CfH_((2f+1−g))F_(g), —OCF₂Cl, —OCF₂—CHFCl,        (C₁-C₂₀)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₂₀)-alkenylcarbonyl, (C₂-C₂₀)-alkynylcarbonyl,        (C₁-C₂₀)-alkoxycarbonyl, (C₆-C₁₂)-aryloxycarbonyl,        (C₇-C₁₆)-aralkoxycarbonyl, (C₃-C₈)-cycloalkoxycarbonyl,        (C₂-C₂₀)-alkenyloxycarbonyl, retinyloxycarbonyl,        (C₂-C₂₀)-alkynyloxycarbonyl, (C₁-C₁₂)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkylcarbonyloxy, (C₆-C₁₂)-arylcarbonyloxy,        (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,        (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy, (C₆-C₁₂)-aryloxycarbonyloxy,        (C₇-C₁₆)-aralkyloxycarbonyloxy, (C₃-C₈)-cycloalkoxycarbonyloxy,        (C₂-C₁₂)-alkenyloxycarbonyloxy, (C₂-C₁₂)-alkynyloxycarbonyloxy,        carbamoyl, N—(C₁-C₁₂)-alkylcarbamoyl,        N,N-di-(C₁-C₁₂)-alkylcarbamoyl, N—(C₃-C₈)-cycloalkylcarbamoyl,        N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,        N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)carbamoyl,        N—(C₁-C₆)-alkyl-N-((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)carbamoyl,        N-(+)-dehydroabietylcarbamoyl,        N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,        N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl,        N—((C₁-C₁₆)-alkoxy-(C₁-C₁₀)-alkyl)carbamoyl,        N—((C₆-C₁₆)-aryloxy-(C₁-C(C₁₀)-alkyl)carbamoyl,        N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₁-C₁)-alkoxy-(C₁-C₁₀)-alkyl)carbamoyl,        N—(C₁-C₁₀)-alkyl-N-((C₆-C₁₂)-aryloxy-(C₁-C₁₀)-alkyl)carbamoyl,        N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyl,        carbamoyloxy, N—(C₁-C₁₂)-alkylcarbamoyloxy,        N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—((C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxyamino,        (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,        (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C₇-C₁₁)-aralkylamino, N-alkyl-aralkylamino,        N-alkyl-arylamino, (C₁-C₁₂)-alkoxyamino,        (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino, (C₁-C₁₂)-alkanoylamino,        (C₃-C₈)-cycloalkanoylamino, (C₆-C₁₂)-aroylamino,        (C₇-C₁₆)-aralkanoylamino,        (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,        (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino, amino-(C₁-C₁₀)-alkyl,        (C₁-C₂₀)-alkylmercapto, (C₁-C₂₀)-alkylsulfinyl,        (C₁-C₂₀)-alkylsulfonyl, (C₆-C₁₂)-arylmercapto,        (C₆-C₁₂)-arylsulfinyl, (C₆-C₁₂)-arylsulfonyl,        (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl,        (C₇-C₁₆)-aralkylsulfonyl, sulfamoyl, N—(C₁-C₁₀)-alkylsulfamoyl,        N,N-di-(C₁-C₁₀)-alkylsulfamoyl, (C₃-C₈)-cycloalkylsulfamoyl,        N—(C₆-C₁₂)-arylsulfamoyl, N—(C₇-C₁₆)-aralkylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,        (C₁-C₁₀)-alkylsulfonamido, (C₇-C₁₆)-aralkylsulfonamido, and        N-((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; where an aryl        radical may be substituted by 1 to 5 substituents selected from        hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₂-C₁₆)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₂-C₁₆)-alkenyl,        (C₂-C₁₂)-alkynyl, (C₁-C₁₆)-alkoxy, (C₁-C₁₆)-alkenyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        —O—[CH₂]_(x)C_(f)H_((2f+1−g))F_(g), —OCF₂Cl, and —OCF₂—CHFCl;

x is 0 to 3;

f is 1 to 8; and

g is 0 or 1 to (2f+1);

including the physiologically active salts and prodrugs derivedtherefrom.

Pyridine-2-carboxamides of Formula I include, but are not limited to,[(3-methoxy-pyridine-2-carbonyl)-amino]-acetic acid,3-methoxypyridine-2-carboxylic acidN-(((hexadecyloxy)-carbonyl)-methyl)-amide hydrochloride,3-methoxypyridine-2-carboxylic acidN-(((1-octyloxy)-carbonyl)-methyl)-amide, 3-methoxypyridine-2-carboxylicacid N-(((hexyloxy)-carbonyl)-methyl)-amide,3-methoxypyridine-2-carboxylic acidN-(((butyloxy)-carbonyl)-methyl)-amide, 3-methoxypyridine-2-carboxylicacid N-(((2-nonyloxy)-carbonyl)-methyl)-amide racemate,3-methoxypyridine-2-carboxylic acidN-(((heptyloxy)-carbonyl)-methyl)-amide,3-benzyloxypyridine-2-carboxylic acidN-(((octyloxy)-carbonyl)-methyl)-amide, 3-benzyloxypyridine-2-carboxylicacid N-(((butyloxy)-carbonyl)-methyl)-amide,5-(((3-(1-butyloxy)-propyl)-amino)-carbonyl)-3-methoxypyridine-2-carboxylicacid N-((benzyloxycarbonyl)-methyl)-amide,5-(((3-(1-butyloxy)-propyl)-amino)-carbonyl)-3-methoxypyridine-2-carboxylicacid N-(((1-butyloxy)-carbonyl)-methyl)-amide,5-(((3-lauryloxy)-propyl)amino)-carbonyl)-3-methoxypyridine-2-carboxylicacid N-(((benzyloxy)-carbonyl)-methyl)-amide,[(3-hydroxy-pyridine-2-carbonyl)-amino]-acetic acid (Compound G), and[(3-methoxy-pyridine-2-carbonyl)-amino]-acetic acid (Compound P).

In certain embodiments, compounds used in the methods of the inventionare quinoline-2-carboxamides. In one embodiment, the compound isselected from a compound of the Formula Ia wherein

-   -   A is —CR⁵R⁶—, and R⁵ and R⁶ are each independently selected from        the group consisting of hydrogen, (C₁-C₆)-alkyl,        (C₃-C₇)-cycloalkyl, aryl, or a substituent of the α-carbon atom        of an α-amino acid, wherein the amino acid is a natural L-amino        acid or its D-isomer;    -   B is —CO₂H or a CO₂-G carboxyl radical, where G is a radical of        an alcohol G-OH in which G is selected from the group consisting        of (C₁-C₂₀)-alkyl radical, (C₃-C₈) cycloalkyl radical,        (C₂-C₂₀)-alkenyl radical, (C₃-C₈)-cycloalkenyl radical, retinyl        radical, (C₂-C₂₀)-alkynyl radical, (C₄-C₂₀)-alkenynyl radical;

X is O;

Q is O;

-   -   R⁴ is selected from the group consisting of hydrogen,        (C₁-C₁₀)-alkyl, (C₂-C₁₀)-alkenyl, (C₂-C₁₀)-alkynyl, wherein        alkenyl or alkynyl contains one or two C—C multiple bonds;        unsubstituted fluoroalkyl radical of the formula        —[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), aryl, heteroaryl, and        (C₇-C₁₁)-aralkyl;    -   R¹, R¹², R¹³, R¹⁴ and R¹⁵ are identical or different and are        selected from the group consisting of hydrogen, hydroxyl,        halogen, cyano, trifluoromethyl, nitro, carboxyl;        (C₁-C₂₀)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₇-C₁₆)-aralkenyl,        (C₇-C₁₆)-aralkynyl, (C₂-C₂₀)-alkenyl, (C₂-C₂₀)-alkynyl,        (C₁-C₂₀)-alkoxy, (C₂-C₂₀)-alkenyloxy, (C₂-C₂₀)-alkynyloxy,        retinyloxy, (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy,        (C₁-C₁₆)-hydroxyalkyl, —O—[CH₂]_(x)CfH_((2f+1−g))F_(g), —OCF₂Cl,        —OCF₂—CHFCl, (C₁-C₂₀)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₂₀)-alkenylcarbonyl, (C₂-C₂₀)-alkynylcarbonyl,        (C₁-C₂₀)-alkoxycarbonyl, (C₆-C₁₂)-aryloxycarbonyl,        (C₇-C₁₆)-aralkoxycarbonyl, (C₃-C₈)-cycloalkoxycarbonyl,        (C₂-C₂₀)-alkenyloxycarbonyl, retinyloxycarbonyl,        (C₂-C₂₀)-alkynyloxycarbonyl, (C₁-C₁₂)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkylcarbonyloxy, (C₆-C₁₂)-arylcarbonyloxy,        (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,        (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy, (C₆-C₁₂)-aryloxycarbonyloxy,        (C₇-C₁₆)-aralkyloxycarbonyloxy, (C₃-C₈)-cycloalkoxycarbonyloxy,        (C₂-C₁₂)-alkenyloxycarbonyloxy, (C₂-C₁₂)-alkynyloxycarbonyloxy,        carbamoyl, N—(C₁-C₁₂)-alkylcarbamoyl,        N,N-di-(C₁-C₁₂)-alkylcarbamoyl, N—(C₃-C₈)-cycloalkylcarbamoyl,        N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,        N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)-carbamoyl,        N-(+)-dehydroabietylcarbamoyl,        N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,        N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl, carbamoyloxy,        N—(C₁-C₁₂)-alkylcarbamoyloxy, N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—((C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxyamino,        (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,        (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C₇-C₁₁)-aralkylamino, N-alkyl-aralkylamino,        N-alkyl-arylamino, (C₁-C₁₂)-alkoxyamino,        (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino, (C₁-C₁₂)-alkanoylamino,        (C₃-C₈)-cycloalkanoylamino, (C₆-C₁₂)-aroylamino,        (C₇-C₁₆)-aralkanoylamino,        (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,        (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino, amino-(C₁-C₁₀)-alkyl,        (C₁-C₂₀)-alkylmercapto, (C₁-C₂₀)-alkylsulfinyl,        (C₁-C₂₀)-alkylsulfonyl, (C₆-C₁₂)-arylmercapto,        (C₆-C₁₂)-arylsulfinyl, (C₆-C₁₂)-arylsulfonyl,        (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl,        (C₇-C₁₆)-aralkylsulfonyl, sulfamoyl, N—(C₁-C₁₀)-alkylsulfamoyl,        N,N-di-(C₁-C₁₀)-alkylsulfamoyl, (C₃-C₈)-cycloalkylsulfamoyl,        N—(C₆-C₁₂)-arylsulfamoyl, N—(C₇-C₁₆)-aralkylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,        (C₁-C₁₀)-alkylsulfonamido, (C₇-C₁₆)-aralkylsulfonamido, and        N-((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; where an aryl        radical may be substituted by 1 to 5 substituents selected from        hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₂-C₁₆)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₂-C₁₆)-alkenyl,        (C₂-C₁₂)-alkynyl, (C₁-C₁₆)-alkoxy, (C₁-C₁₆)-alkenyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        —O—[CH₂]_(x)C_(f)H_((2f+1−g))F_(g), —OCF₂Cl, and —OCF₂—CHFCl;

x is 0 to 3;

f is 1 to 8; and

g is 0 or 1 to (2f+1);

including the physiologically active salts and prodrugs derivedtherefrom.

Quinoline-2-carboxamides of Formula Ia include, but are not limited to,N-((3-Hydroxy-6-isopropoxy-quinoline-2-carbonyl)-amino)-acetic acid(Compound H),N-((6-(1-butyloxy)-3-hydroxyquinolin-2-yl)-carbonyl)-glycine,[(3-hydroxy-6-trifluoromethoxy-quinoline-2-carbonyl)-amino]-acetic acid(Compound I), N-((7-chloro-3-hydroxyquinolin-2-yl)-carbonyl)-glycine,and [(6-chloro-3-hydroxy-quinoline-2-carbonyl)-amino]-acetic acid(Compound O).

In certain embodiments, compounds used in the methods of the inventionare isoquinoline-3-carboxamides. In one embodiment, the compound isselected from a compound of the Formula Ib wherein

-   -   A is —CR⁵R⁶—, and R⁵ and R⁶ are each independently selected from        the group consisting of hydrogen, (C₁-C₆)-alkyl,        (C₃-C₇)-cycloalkyl, aryl, or a substituent of the α-carbon atom        of an α-amino acid, wherein the amino acid is a natural L-amino        acid or its D-isomer;    -   B is —CO₂H or a CO₂-G carboxyl radical, where G is a radical of        an alcohol G-OH in which G is selected from the group consisting        of (C₁-C₂₀)-alkyl radical, (C₃-C₈) cycloalkyl radical,        (C₂-C₂₀)-alkenyl radical, (C₃-C₈)-cycloalkenyl radical, retinyl        radical, (C₂-C₂₀)-alkynyl radical, (C₄-C₂₀)-alkenynyl radical;

X is O;

Q is O;

-   -   R⁴ is selected from the group consisting of hydrogen,        (C₁-C₁₀)-alkyl, (C₂-C₁₀)-alkenyl, (C₂-C₁₀)-alkynyl, wherein        alkenyl or alkynyl contains one or two C—C multiple bonds;        unsubstituted fluoroalkyl radical of the formula        —[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), aryl, heteroaryl, and        (C₇-C₁₁)-aralkyl;    -   R³, R¹⁶, R¹⁷, R¹⁸ and R¹⁹ are identical or different and are        selected from the group consisting of hydrogen, hydroxyl,        halogen, cyano, trifluoromethyl, nitro, carboxyl;        (C₁-C₂₀)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₇-C₁₆)-aralkenyl,        (C₇-C₁₆)-aralkynyl, (C₂-C₂₀)-alkenyl, (C₂-C₂₀)-alkynyl,        (C₁-C₂₀)-alkoxy, (C₂-C₂₀)-alkenyloxy, (C₂-C₂₀)-alkynyloxy,        retinyloxy, (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy,        (C₁-C₁₆)-hydroxyalkyl, —O—[CH₂]_(x)CfH_((2f+1−g))F_(g), —OCF₂Cl,        —OCF₂—CHFCl, (C₁-C₂₀)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₂₀)-alkenylcarbonyl, (C₂-C₂₀)-alkynylcarbonyl,        (C₁-C₂₀)-alkoxycarbonyl, (C₆-C₁₂)-aryloxycarbonyl,        (C₇-C₁₆)-aralkoxycarbonyl, (C₃-C₈)-cycloalkoxycarbonyl,        (C₂-C₂₀)-alkenyloxycarbonyl, retinyloxycarbonyl,        (C₂-C₂₀)-alkynyloxycarbonyl, (C₁-C₁₂)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkylcarbonyloxy, (C₆-C₁₂)-arylcarbonyloxy,        (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,        (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy, (C₆-C₁₂)-aryloxycarbonyloxy,        (C₇-C₁₆)-aralkyloxycarbonyloxy, (C₃-C₈)-cycloalkoxycarbonyloxy,        (C₂-C₁₂)-alkenyloxycarbonyloxy, (C₂-C₁₂)-alkynyloxycarbonyloxy,        carbamoyl, N—(C₁-C₁₂)-alkylcarbamoyl,        N,N-di-(C₁-C₁₂)-alkylcarbamoyl, N—(C₃-C₈)-cycloalkylcarbamoyl,        N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,        N—((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)-carbamoyl,        N-(+)-dehydroabietylcarbamoyl,        N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,        N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl, carbamoyloxy,        N—(C₁-C₁₂)-alkylcarbamoyloxy, N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N-((C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N-((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxyamino,        (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,        (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C₇-C₁₁)-aralkylamino, N-alkyl-aralkylamino,        N-alkyl-arylamino, (C₁-C₁₂)-alkoxyamino,        (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino, (C₁-C₁₂)-alkanoylamino,        (C₃-C₈)-cycloalkanoylamino, (C₆-C₁₂)-aroylamino,        (C₇-C₁₆)-aralkanoylamino,        (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,        (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino, amino-(C₁-C₁₀)-alkyl,        (C₁-C₂₀)-alkylmercapto, (C₁-C₂₀)-alkylsulfinyl,        (C₁-C₂₀)-alkylsulfonyl, (C₆-C₁₂)-arylmercapto,        (C₆-C₁₂)-arylsulfinyl, (C₆-C₁₂)-arylsulfonyl,        (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl,        (C₇-C₁₆)-aralkylsulfonyl, sulfamoyl, N—(C₁-C₁₀)-alkylsulfamoyl,        N,N-di-(C₁-C₁₀)-alkylsulfamoyl, (C₃-C₈)-cycloalkylsulfamoyl,        N—(C₆-C₁₂)-arylsulfamoyl, N—(C₇-C₁₆)-aralkylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,        (C₁-C₁₀)-alkylsulfonamido, (C₇-C₁₆)-aralkylsulfonamido, and        N-((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; where an aryl        radical may be substituted by 1 to 5 substituents selected from        hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₂-C₁₆)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₂-C₁₆)-alkenyl,        (C₂-C₁₂)-alkynyl, (C₁-C₁₆)-alkoxy, (C₁-C₁₆)-alkenyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        —O—[CH₂]_(x)C_(f)H_((2f+1−g))F_(g), —OCF₂Cl, and —OCF₂—CHFCl;

x is 0 to 3;

f is 1 to 8; and

g is 0 or 1 to (2f+1);

including the physiologically active salts and prodrugs derivedtherefrom.

Isoquinoline-3-carboxamides of Formula Ib include, but are not limitedto,N-((1-chloro-4-hydroxy-7-(2-propyloxy)isoquinolin-3-yl)-carbonyl)-glycine,N-((1-chloro-4-hydroxy-6-(2-propyloxy)isoquinolin-3-yl)-carbonyl)-glycine,N-((1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-acetic acid(Compound B),N-((1-chloro-4-hydroxy-7-methoxyisoquinolin-3-yl)-carbonyl)-glycine,N-((1-chloro-4-hydroxy-6-methoxyisoquinolin-3-yl)-carbonyl)-glycine,N-((7-butyloxy)-1-chloro-4-hydroxyisoquinolin-3-yl)-carbonyl)-glycine,N-((6-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid (Compound J),((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid methyl ester (Compound K),N-((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid (Compound L),N-((8-chloro-4-hydroxyisoquinolin-3-yl)-carbonyl)-glycine,N-((7-butoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino)-acetic acid (M),[(1,7-dichloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid(Compound R),{[4-hydroxy-1-(naphthalen-2-yloxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-1-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-1-(3-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-(3-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-(4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-(2-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-1-(2-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid, [(4-hydroxy-1-phenylamino-isoquinoline-3-carbonyl)-amino]-aceticacid, [(1-chloro-4-methoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-chloro-4-ethoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-1-methoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-ethoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-1-phenyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-1-methyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-1-methoxymethyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-dimethylcarbamoyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-hydroxy-1-methyl-6-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid (Compound U),[(4-benzyloxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-ethoxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-dimethylcarbamoyl-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-hydroxy-1-methoxymethyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-1-p-tolyl-isoquinoline-3-carbonyl)-amino]-acetic acid,{[7-(4-fluoro-phenoxy)-4-hydroxy-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid (Compound T),{[1-chloro-4-hydroxy-6-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-6-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-4-hydroxy-7-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-7-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-4-hydroxy-6-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-6-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-7-(4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[7-(4-fluoro-phenoxy)4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-6-(4-fluoro-phenoxy)4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,[(7-benzenesulfinyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(7-benzenesulfonyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(6-benzenesulfinyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(6-benzenesulfonyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,{[4-hydroxy-7-(4-methoxy-benzenesulfonylamino)-isoquinoline-3-carbonyl]-amino}-aceticacid,[(4-hydroxy-1-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,{[1-(4-chloro-phenylsulfanyl)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid, [(4-1-p-tolylsulfanyl-isoquinoline-3-carbonyl)-amino]-acetic acid,{[4-hydroxy-1-(3-methoxy-phenylsulfanyl)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-1-(2-methoxy-phenylsulfanyl)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-1-(naphthalen-2-ylsulfanyl)-isoquinoline-3-carbonyl]-amino}-aceticacid,[(1-benzenesulfinyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-benzenesulfonyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-6,7-diphenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-6,7-diphenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,{[4-hydroxy-7-(4-nitro-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid, [(4-mercapto-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-mercapto-7-trifluoromethyl-isoquinoline-3-carbonyl)-amino]-aceticacid,{[7-(4-chloro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(4-chloro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(3-fluoro-5-methoxy-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[7-(3-fluoro-5-methoxy-phenoxy)4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[7-(3,4-difluoro-phenoxy)4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(3,4-difluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-7-(4-trifluoromethoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-6-(4-trifluoromethoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,2-(s)-{[7-(4-chloro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-{[6-(4-chloro-phenoxy)4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,2-{[7-(3,4-difluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-[(4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-propionicacid.,2-(r)-[(4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-propionicacid,2-(r)-[(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-propionicacid,2-(s)-{[4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-[(7-benzenesulfonyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(r)-2-[(4-hydroxy-1-methoxymethyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(4-hydroxy-1-methoxymethyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(4-mercapto-7-phenoxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-{[1-(4-chloro-phenylsulfanyl)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,(r)-2-{[1-(4-chloro-phenylsulfanyl)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,[(4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-hydroxy-6-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-6-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-6-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid(Compound S),[(4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-chloro-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,{[7-(2,6-dimethyl-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-chloro-7-(2,6-dimethyl-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[1-bromo-7-(2,6-dimethyl-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,[(1-bromo-7-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-6-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-7-trifluoromethyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-6-trifluoromethyl-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-1-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1,7-dibromo-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(7-bromo-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(6-bromo-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-bromo-7-fluoro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(7-fluoro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-chloro-7-fluoro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(1-bromo-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-6-phenyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-7-phenyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-chloro-4-hydroxy-6-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-7-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-6-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-7-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-5-phenyl-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-8-phenyl-isoquinoline-3-carbonyl)-amino]-acetic acid ,[(1-chloro-4-hydroxy-5-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-chloro-4-hydroxy-8-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-5-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-4-hydroxy-8-phenyl-isoquinoline-3-carbonyl)-amino]-aceticacid, [(1-ethylsulfanyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,{[4-hydroxy-1-(4-methoxy-phenylsulfanyl)-isoquinoline-3-carbonyl]-amino}-aceticacid, [(1-chloro-4-hydroxy-7-iodo-isoquinoline-3-carbonyl)-amino]-aceticacid, [(1-chloro-4-hydroxy-6-iodo-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-7-iodo-isoquinoline-3-carbonyl)-amino]-acetic acid,[(1-bromo-4-hydroxy-7-methyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-7-butoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-bromo-6-butoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,(r)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,(s)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-hydroxy-propionicacid,2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-2-methyl-propionicacid,2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-2-methyl-propionicacid,(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-(1h-imidazol-4-yl)-propionicacid (trifluoro-acetic acid salt),(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-(1h-imidazol-4-yl)-propionicacid (trifluoro-acetic acid salt),(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(r)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(s)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(s)-2-[(6-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-methyl-butyricacid,(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(r)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(s)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-phenyl-propionicacid,(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-(4-hydroxy-phenyl)-propionicacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-3-(4-hydroxy-phenyl)-propionicacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino}-3-(4-hydroxy-phenyl)-propionicacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinolie-3-carbonyl)-amino]-3-(4-hydroxy-phenyl)-propionicacid,(r)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-(4-hydroxy-phenyl)-propionicacid,(s)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-3-(4-hydroxy-phenyl)-propionicacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-pentanoicacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-pentanoicacid,(r)-1-(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-pyrrolidine-2-carboxylicacid,(s)-1-(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-pyrrolidine-2-carboxylicacid,(r)-1-(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-pyrrolidine-2-carboxylicacid,(s)-1-(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-pyrrolidine-2-carboxylicacid,(r)-6-amino-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid (trifluoro-acetic acid salt),(s)-6-amino-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid (trifluoro-acetic acid salt),(r)-6-amino-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid, trifluoroacetic acid salt,(s)-6-amino-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid (trifluoro-acetic acid salt),(r)-6-amino-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid, trifluoroacetic acid salt,(s)-6-amino-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-hexanoicacid (trifluoro-acetic acid salt),(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-succinicacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-succinicacid,(r)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-succinicacid,(s)-2-[(1-chloro-4-hydroxy-6-isopropoxy-isoquinoline-3-carbonyl)-amino]-succinicacid,(r)-2-[(1-chloro-4-hydroxy-7-isopropoxy-isoquinoline-3-carbonyl)-amino]-succinicacid,(r)-2-[(6-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(r)-2-[(7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(r)-2-[(1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(6-isopropoxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(r)-2-[6-isopropoxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,(s)-2-[(7-isopropoxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino-propionicacid, (r)-2-[(7-isopropoxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]propionic acid,{[7-(3,5-difluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-acetic acid,{[6-(3,5-difluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,({7-[4-(4-fluoro-phenoxy)-phenoxy]4-hydroxy-isoquinoline-3-carbonyl}-amino)-aceticacid,({6-[4-(4-fluoro-phenoxy)-phenoxy]-4-hydroxy-isoquinoline-3-carbonyl}-amino)-aceticacid,{[7-(3-chloro-4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(3-chloro-4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-aceticacid,(s)-2-{[7-(3-fluoro-5-methoxy-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-[(7-cyclohexyloxy-4-hydroxy-isoquinoline-3-carbonyl)-amino]-propionicacid,2-(s)-{[7-(4-fluoro-phenoxy)-4-hydroxy-1-methyl-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-{[7-(4-fluoro-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino}-propionicacid,2-(s)-[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-propionicacid,2-(s)-[(4-hydroxy-1-methyl-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-propionicacid,2-(s)-{[4-hydroxy-7-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-propionicacid,{[7-(4-chloro-phenoxy)-4-hydroxy-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid,{[6-(4-chloro-phenoxy)-4-hydroxy-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid,{[7-(3,5-difluoro-phenoxy)-4-hydroxy-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-7-(4-methoxy-phenoxy)-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid,{[4-hydroxy-6-(4-methoxy-phenoxy)-1-methyl-isoquinoline-3-carbonyl]-amino}-aceticacid, [(6-cyclohexyloxy-4-hydr6xy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(7-cyclohexyloxy-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(7-cyclohexyloxy-4-hydroxy-1-methyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(7-cyclohexylsulfanyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(7-cyclohexanesulfonyl-4-hydroxy-isoquinoline-3-carbonyl)-amino]-aceticacid, [(4-hydroxy-1-isobutyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-ethyl-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid,[(1-dimethylaminomethyl-4-hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,[(4-hydroxy-1-methyl-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-aceticacid,{[4-hydroxy-1-methyl-7-(4-trifluoromethyl-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid.

In other embodiments, compounds used in the methods of the invention areselected from a compound of the Formula II

where

-   -   R²⁸ is hydrogen, nitro, amino, cyano, halogen, (C₁-C₄)-alkyl,        carboxy or a metabolically labile ester derivative thereof;        (C₁-C₄)-alkylamino, di-(C₁-C₄)-alkylamino,        (C₁-C₆)-alkoxycarbonyl, (C₂-C₄)-alkanoyl, hydroxy-(C₁-C₄)-alkyl,        carbamoyl, N—(C₁-C₄)-alkylcarbamoyl, (C₁-C₄)-alkylthio,        (C₁-C₄)-alkylsulfinyl, (C₁-C₄)-alkylsulfonyl, phenylthio,        phenylsulfinyl, phenylsulfonyl, said phenyl or phenyl groups        being optionally substituted with 1 to 4 identical or different        halogen, (C₁-C₄)-alkyoxy, (C₁-C₄)-alkyl, cyano, hydroxy,        trifluoromethyl, fluoro-(C₁-C₄)-alkylthio,        fluoro-(C₁-C₄)-alkylsulfinyl, fluoro-(C₁-C₄)-alkylsulfonyl,        (C₁-C₄)-alkoxy-(C₂-C₄)-alkoxycarbonyl,        N,N-di-[(C₁-C₄)-alkyl]carbamoyl-(C₁-C₄)-alkoxycarbonyl,        (C₁-C₄)-alkylamino-(C₂-C₄)-alkoxycarbonyl,        di-(C₁-C₄)-alkylamino-(C₂-C₄)-alkoxycarbonyl,        (C₁-C₄)-alkoxy-(C₂-C₄)-alkoxy-(C₂-C₄)-alkoxycarbonyl,        (C₂-C₄)-alkanoyloxy-C₁-C₄)-alkyl, or        N-[amino-(C₂-C₈)-alkyl]-carbamoyl;    -   R²⁹ is hydrogen, hydroxy, amino, cyano, halogen, (C₁-C₄)-alkyl,        carboxy or metabolically labile ester derivative thereof,        (C₁-C₄)-alkylamino, di-(C₁-C₄)-alkylamino,        (C₁-C₆)-alkoxycarbonyl, (C₂-C₄)-alkanoyl, (C₁-C₄)-alkoxy,        carboxy-(C₁-C₄)-alkoxy, (C₁-C₄)-alkoxycarbonyl-(C₁-C₄)-alkoxy,        carbamoyl, N—(C₁-C₈)-alkylcarbamoyl,        N,N-di-(C₁-C₈)-alkylcarbamoyl,        N-[amino-(C₂-C₈)-alkyl]-carbamoyl,        N-[(C₁-C₄)-alkylamino-(C₁-C₈)-alkyl]-carbamoyl,        N-[di-(C₁-C₄)-alkylamino-(C₁-C₈)-alkyl]]-carbamoyl,        N-cyclohexylcarbamoyl, N-[cyclopentyl]-carbamoyl,        N—(C₁-C₄)-alkylcyclohexylcarbamoyl,        N—(C₁-C₄)-alkylcyclopentylcarbamoyl, N-phenylcarbamoyl,        N—(C₁-C₄)-alkyl-N-phenylcarbamoyl, N,N-diphenylcarbamoyl,        N-[phenyl-(C₁-C₄)-alkyl]-carbamoyl,        N—(C₁-C₄)-alkyl-N-[phenyl-(C₁-C₄)-alkyl]-carbamoyl, or        N,N-di-[phenyl-(C₁-C₄)-alkyl]-carbamoyl, said phenyl or phenyl        groups being optionally substituted with 1 to 4 identical or        different halogen, (C₁-C₄)-alkyoxy, (C₁-C₄)-alkyl, cyano,        hydroxy, trifluoromethyl, N-[(C₂-C₄)-alkanoyl]-carbamoyl,        N-[(C₁-C₄)-alkoxycarbonyl]-carbamoyl,        N-[fluoro-(C₂-C₆)-alkyl]-carbamoyl,        N,N-[fluoro-(C₂-C₆)-alkyl]-N—(C₁-C₄)-alkylcarbamoyl,        N,N-[di-fluoro-(C₂-C₆)-alkyl]carbamoyl, pyrrolidin-1-ylcarbonyl,        piperidinocarbonyl, piperazin-1-ylcarbonyl, morpholinocarbonyl,        wherein the heterocyclic group, is optionally substituted with 1        to 4, (C₁-C₄)-alkyl, benzyl,        1,2,3,4-tetrahydro-isoquinolin-2-ylcarbonyl,        N,N-[di-(C₁-C₄)-alkyl]-thiocarbamoyl, N—(C₂-C₄)-alkanoylamino,        or N-[(C₁-C₄)-alkoxycarbonyl]-amino;    -   R³⁰ is hydrogen, (C₁-C₄)-alkyl, (C₂-C₄)-alkoxy, halo, nitro,        hydroxy, fluoro-(1-4C)alkyl, or pyridinyl;    -   R³¹ is hydrogen, (C₁-C₄)-alkyl, (C₂-C₄)-alkoxy, halo, nitro,        hydroxy, fluoro-(C₁-C₄)-alkyl, pyridinyl, or methoxy;    -   R³² is hydrogen, hydroxy, amino, (C₁-C₄)-alkylamino,        di-(C₁-C₄)-alkylamino, halo, (C₁-C₄)-alkoxy-(C₂-C₄)-alkoxy,        fluoro-(C₁-C₆)-alkoxy, pyrrolidin-1-yl, piperidino,        piperazin-1-yl, or morpholino, wherein the heterocyclic group is        optionally substituted with 1 to 4 identical or different        (C₁-C₄)-alkyl or benzyl; and    -   R³³ and R³⁴ are individually selected from hydrogen,        (C₁-C₄)-alkyl, and (C₁-C₄)-alkoxy; including        pharmaceutically-acceptable salts and pro-drugs derived        therefrom.

Exemplary compounds of Formula II are described in U.S. Pat. Nos.5,916,898 and 6,200,974, and International Publication No. WO 99/21860.All compounds listed in the foregoing patents and publication, inparticular, those listed in the compound claims and the final productsof the working examples, are hereby incorporated into the presentapplication by reference herein. Exemplary compounds of Formula (II)include 4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylic acid(Compound A) (see, e.g., Seki et al. (1974) Chem Abstracts 81:424, No.21), 3-carboxy-5-hydroxy-4-oxo-3,4-dihydro-1,10-phenanthroline,3-carboxy-5-methoxy-4-oxo-3,4-dihydro-1,10-phenanthroline,5-methoxy-4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylic acid ethylester, 5-methoxy-4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylicacid (Compound Q), and3-carboxy-8-hydroxy-4-oxo-3,4-dihydro-1,10-phenanthroline.

In other embodiments, compounds used in the methods of the invention areselected from a compound of the Formula III

or pharmaceutically acceptable salts thereof, wherein:

a is an integer from 1 to 4;

b is an integer from 0 to 4;

c is an integer from 0 to 4;

-   -   Z is selected from the group consisting of (C₃-C₁₀) cycloalkyl,        (C₃-C₁₀) cycloalkyl independently substituted with one or more        Y¹, 3-10 membered heterocycloalkyl and 3-10 membered        heterocycloalkyl independently substituted with one or more Y¹;        (C₅-C₂₀) aryl, (C₅-C₂₀) aryl independently substituted with one        or more Y¹, 5-20 membered heteroaryl and 5-20 membered        heteroaryl independently substituted with one or more Y¹;    -   Ar¹ is selected from the group consisting of (C₅-C₂₀) aryl,        (C₅-C₂₀) aryl independently substituted with one or more Y²,        5-20 membered heteroaryl and 5-20 membered heteroaryl        independently substituted with one or more Y²;    -   each Y¹ is independently selected from the group consisting of a        lipophilic functional group, (C₅-C₂₀) aryl, (C₆-C₂₆) alkaryl,        5-20 membered heteroaryl and 6-26 membered alk-heteroaryl;    -   each Y² is independently selected from the group consisting of        −R′, —OR′, —OR″, —SR′, —SR″, —NR′R′, —NO₂, —CN, -halogen,        -trihalomethyl, trihalomethoxy, —C(O)R′, —C(O)OR′, —C(O)NR′R′,        —C(O)NR′OR′, —C(NR′R′)═NOR′, —NR′—C(O)R′, —SO₂R′, —SO₂R″,        —NR′—SO₂—R′, —NR′—C(O)—NR′R′, tetrazol-5-yl, —NR′—C(O)—OR′,        —C(NR′R′)═NR′, —S(O)—R′, —S(O)—R″, and —NR′—C(S)—NR′R′; and    -   each R′ is independently selected from the group consisting of        —H, (C₁-C₈) alkyl, (C₂-C₈) alkenyl, and (C₂-C₈) alkynyl; and    -   each R″ is independently selected from the group consisting of        (C₅-C₂₀) aryl and (C₅-C₂₀) aryl independently substituted with        one or more —OR′, —SR′, —NR′R′, —NO₂, —CN, halogen or        trihalomethyl groups,

-   or wherein c is 0 and Ar¹ is an N′ substituted urea-aryl, the    compound has the structural Formula IIIa:

or pharmaceutically acceptable salts thereof, wherein:

a, b, and Z are as defined above; and

R³⁵ and R³⁶ are each independently selected from the group consisting ofhydrogen, (C₁-C₈) alkyl, (C₂-C₈) alkenyl, (C₂-C₈) alkynyl, (C₃-C₁₀)cycloalkyl, (C₅-C₂₀) aryl, (C₅-C₂₀) substituted aryl, (C₆-C₂₆) alkaryl,(C₆-C₂₆) substituted alkaryl, 5-20 membered heteroaryl, 5-20 memberedsubstituted heteroaryl, 6-26 membered alk-heteroaryl, and 6-26 memberedsubstituted alk-heteroaryl; and

-   -   R³⁷ is independently selected from the group consisting of        hydrogen, (C₁-C₈) alkyl, (C₂-C₈) alkenyl, and (C₂-C₈) alkynyl.

Exemplary compounds of Formula (III) are described in InternationalPublication No. WO 00/50390. All compounds listed in WO 00/50390, inparticular, those listed in the compound claims and the final productsof the working examples, are hereby incorporated into the presentapplication by reference herein. Exemplary compounds of Formula (III)include3-{[4-(3,3-dibenzyl-ureido)-benzenesulfonyl]-[2-(4-methoxy-phenyl)-ethyl]-amino}-N-hydroxy-propionamide(Compound C),3-{{4-[3-(4-chloro-phenyl)-ureido]-benzenesulfonyl}-[2-(4-methoxy-phenyl)-ethyl]-amino}-N-hydroxy-propionamide,and3-{{4-[3-(1,2-diphenyl-ethyl)-ureido]-benzenesulfonyl}-[2-(4-methoxy-phenyl)-ethyl]-amino}-N-hydroxy-propionamide.

In certain embodiments, compounds used in the methods of the inventionare selected from a compound of the formula (IV)

wherein

-   -   R¹ are selected from the group consisting of hydrogen,        (C₁-C₆)-alkyl, (C₃-C₇)-cycloalkyl, aryl, or a substituent of the        α-carbon atom of an α-amino acid, wherein the amino acid is a        natural L-amino acid or its D-isomer;    -   B is —CO₂H or a CO₂-G carboxyl radical, where G is a radical of        an alcohol G-OH in which G is selected from the group consisting        of (C₁-C₂₀)-alkyl radical, (C₃-C₈) cycloalkyl radical,        (C₂-C₂₀)-alkenyl radical, (C₃-C₈)-cycloalkenyl radical, retinyl        radical, (C₂-C₂₀)-alkynyl radical, (C₄-C₂₀)-alkenynyl radical;    -   R² is selected from the group consisting of hydrogen,        (C₁-C₁₀)-alkyl, (C₂-C₁₀)-alkenyl, (C₂-C₁₀)-alkynyl, wherein        alkenyl or alkynyl contains one or two C—C multiple bonds;

unsubstituted fluoroalkyl radical of the formula—[CH₂]_(x)—C_(f)H_((2f+1−g))—F_(g), aryl, heteroaryl, and(C₇-C₁₁)-aralkyl;

one of D or M is —S—, and the other is ═C(R⁵)—;

-   -   R³, R⁴, and R⁵ are identical or different and are selected from        the group consisting of hydrogen, hydroxyl, halogen, cyano,        trifluoromethyl, nitro, carboxyl; (C₁-C₂₀)-alkyl,        (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy, (C₆-C₁₂)-aryl,        (C₇-C₁₆)-aralkyl, (C₇-C₁₆)-aralkenyl, (C₇-C₁₆)-aralkynyl,        (C₂-C₂₀)-alkenyl, (C₂-C₂₀)-alkynyl, (C₁-C₂₀)-alkoxy,        (C₂-C₂₀)-alkenyloxy, (C₂-C₂₀)-alkynyloxy, retinyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₁₆)-hydroxyalkyl,        —O—[CH₂]_(x)CfH_((2f+1−g))F_(g), —OCF₂Cl, —OCF₂—CHFCl,        (C₁-C₂₀)-alkylcarbonyl, (C₃-C₈)-cycloalkylcarbonyl,        (C₆-C₁₂)-arylcarbonyl, (C₇-C₁₆)-aralkylcarbonyl, cinnamoyl,        (C₂-C₂₀)-alkenylcarbonyl, (C₂-C₂₀)-alkynylcarbonyl,        (C₁-C₂₀)-alkoxycarbonyl, (C₆-C₁₂)-aryloxycarbonyl,        (C₇-C₁₆)-aralkoxycarbonyl, (C₃-C₈)-cycloalkoxycarbonyl,        (C₂-C₂₀)-alkenyloxycarbonyl, retinyloxycarbonyl,        (C₂-C₂₀)-alkynyloxycarbonyl, (C₁-C₁₂)-alkylcarbonyloxy,        (C₃-C₈)-cycloalkylcarbonyloxy, (C₆-C₁₂)-arylcarbonyloxy,        (C₇-C₁₆)-aralkylcarbonyloxy, cinnamoyloxy,        (C₂-C₁₂)-alkenylcarbonyloxy, (C₂-C₁₂)-alkynylcarbonyloxy,        (C₁-C₁₂)-alkoxycarbonyloxy, (C₆-C₁₂)-aryloxycarbonyloxy,        (C₇-C₁₆)-aralkyloxycarbonyloxy, (C₃-C₈)-cycloalkoxycarbonyloxy,        (C₂-C₁₂)-alkenyloxycarbonyloxy, (C₂-C₁₂)-alkynyloxycarbonyloxy,        carbamoyl, N—(C₁-C₁₂)-alkylcarbamoyl,        N,N-di-(C₁-C₁₂)-alkylcarbamoyl, N—(C₃-C₈)-cycloalkylcarbamoyl,        N,N-dicyclo-(C₃-C₈)-alkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₃-C₈)-cycloalkylcarbamoyl,        N-((C₃-C₈)-cycloalkyl-(C₁-C₆)-alkyl)-carbamoyl,        N-(+)-dehydroabietylcarbamoyl,        N—(C₁-C₆)-alkyl-N-(+)-dehydroabietylcarbamoyl,        N—(C₆-C₁₂)-arylcarbamoyl, N—(C₇-C₁₆)-aralkylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₆)-arylcarbamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyl, carbamoyloxy,        N—(C₁-C₁₂)-alkylcarbamoyloxy, N,N-di-(C₁-C₁₂)-alkylcarbamoyloxy,        N—(C₃-C₈)-cycloalkylcarbamoyloxy, N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylcarbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylcarbamoyloxy,        N—((C₁-C₁₀)-alkyl)-carbamoyloxy,        N—(C₁-C₁₀)-alkyl-N—((C₇-C₁₆)-aralkyloxy-(C₁-C₁₀)-alkyl)-carbamoyloxyamino,        (C₁-C₁₂)-alkylamino, di-(C₁-C₁₂)-alkylamino,        (C₃-C₈)-cycloalkylamino, (C₃-C₁₂)-alkenylamino,        (C₃-C₁₂)-alkynylamino, N—(C₆-C₁₂)-arylamino,        N—(C₇-C₁₁)-aralkylamino, N-alkyl-aralkylamino,        N-alkyl-arylamino, (C₁-C₁₂)-alkoxyamino,        (C₁-C₁₂)-alkoxy-N—(C₁-C₁₀)-alkylamino, (C₁-C₁₂)-alkanoylamino,        (C₃-C₈)-cycloalkanoylamino, (C₆-C₁₂)-aroylamino,        (C₇-C₆)-aralkanoylamino,        (C₁-C₁₂)-alkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₃-C₈)-cycloalkanoyl-N—(C₁-C₁₀)-alkylamino,        (C₆-C₁₂)-aroyl-N—(C₁-C₁₀)-alkylamino,        (C₇-C₁₁)-aralkanoyl-N—(C₁-C₁₀)-alkylamino, amino-(C₁-C₁₀)-alkyl,        (C₁-C₂₀)-alkylmercapto, (C₁-C₂₀)-alkylsulfinyl,        (C₁-C₂₀)-alkylsulfonyl, (C₆-C₁₂)-arylmercapto,        (C₆-C₁₂)-arylsulfinyl, (C₆-C₁₂)-arylsulfonyl,        (C₇-C₁₆)-aralkylmercapto, (C₇-C₁₆)-aralkylsulfinyl,        (C₇-C₁₆)-aralkylsulfonyl, sulfamoyl, N—(C₁-C₁₀)-alkylsulfamoyl,        N,N-di-(C₁-C₁₀)-alkylsulfamoyl, (C₃-C₈)-cycloalkylsulfamoyl,        N—(C₆-C₁₂)-arylsulfamoyl, N—(C₇-C₁₆)-aralkylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₆-C₁₂)-arylsulfamoyl,        N—(C₁-C₁₀)-alkyl-N—(C₇-C₁₆)-aralkylsulfamoyl,        (C₁-C₁₀)-alkylsulfonamido, (C₇-C₁₆)-aralkylsulfonamido, and        N-((C₁-C₁₀)-alkyl-(C₇-C₁₆)-aralkylsulfonamido; where an aryl        radical may be substituted by 1 to 5 substituents selected from        hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl,        (C₂-C₁₆)-alkyl, (C₃-C₈)-cycloalkyl, (C₃-C₈)-cycloalkoxy,        (C₆-C₁₂)-aryl, (C₇-C₁₆)-aralkyl, (C₂-C₁₆)-alkenyl,        (C₂-C₁₂)-alkynyl, (C₁-C₁₆)-alkoxy, (C₁-C₁₆)-alkenyloxy,        (C₆-C₁₂)-aryloxy, (C₇-C₁₆)-aralkyloxy, (C₁-C₈)-hydroxyalkyl,        —O—[CH₂]_(x)C_(f)H_((2f+1−g))F_(g), —OCF₂Cl, and —OCF₂—CHFCl;

x is 0 to 3;

f is 1 to 8; and

g is 0 or 1 to (2f+1);

including the physiologically active salts and prodrugs derivedtherefrom.

Compounds of Formula (IV) include, but are not limited to,[(2-bromo-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(2-bromo-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,{[4-hydroxy-2-(4-methoxy-phenyl)-thieno[2,3-c]pyridine-S-carbonyl]-amino}-aceticacid,{[7-hydroxy-2-(4-methoxy-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,[(4-hydroxy-2,7-dimethyl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(7-hydroxy-2,4-dimethyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,{[7-hydroxy-4-methyl-2-(4-phenoxy-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[4-hydroxy-2-(4-phenoxy-phenyl)-7-methyl-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[4-hydroxy-2-(4-phenoxy-phenyl)-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[7-hydroxy-2-(4-phenoxy-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,[(2,7-dibromo-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(2-bromo-7-chloro-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid, [(7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-acetic acid,[(4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-acetic acid,[(2-bromo-4-chloro-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(2,4-dibromo-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-hydroxy-2-phenylsulfanyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(4-hydroxy-2-phenylsulfanyl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(4-hydroxy-2,7-diphenyl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(7-hydroxy-2,4-diphenyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-hydroxy-2-styryl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-hydroxy-2-phenoxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-hydroxy-2-phenethyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[7-hydroxy-2-(3-trifluoromethyl-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[4-bromo-7-hydroxy-2-(3-trifluoromethyl-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[4-cyano-7-hydroxy-2-(3-trifluoromethyl-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,[(2-cyano-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,{[7-hydroxy-2-(4-trifluoromethyl-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[7-hydroxy-2-(2-trifluoromethyl-phenyl)-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[4-bromo-3-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[3-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[3-(4-fluoro-phenyl)-7-hydroxy-4-methyl-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[4-cyano-3-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[2-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[2-(4-fluoro-phenyl)-7-hydroxy-4-methyl-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[2,3-bis-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,{[7-bromo-3-(4-fluoro-phenyl)-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[3-(4-fluoro-phenyl)-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[2-(4-fluoro-phenyl)-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[2-(4-fluoro-phenyl)-4-hydroxy-7-methyl-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,[(7-chloro-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(4-chloro-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-bromo-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(4-bromo-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(4-hydroxy-7-phenyl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(7-hydroxy-4-phenyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,{[7-(4-fluoro-phenyl)-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl]-amino}-aceticacid,{[4-(4-fluoro-phenyl)-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl]-amino}-aceticacid,2-(7-(furan-2-yl)-4-hydroxythieno[2,3-c]pyridine-5-carboxamido)aceticacid,[(4-furan-2-yl-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-furan-3-yl-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(4-furan-3-yl-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,2-(4-hydroxy-7-(thiophen-2-yl)thieno[2,3-c]pyridine-5-carboxamido)aceticacid,[(7-hydroxy-4-thiophen-2-yl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(4-hydroxy-7-thiophen-3-yl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(7-hydroxy-4-thiophen-3-yl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(4-hydroxy-7-methyl-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(7-hydroxy-4-methyl-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-ethynyl-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid,[(4-ethynyl-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid,[(7-cyano-4-hydroxy-thieno[2,3-c]pyridine-5-carbonyl)-amino]-aceticacid, and[(4-cyano-7-hydroxy-thieno[3,2-c]pyridine-6-carbonyl)-amino]-aceticacid.

Exemplary compounds for use in the present methods include4-oxo-1,4-dihydro-[1,10]phenanthroline-3-carboxylic acid,N-((1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-acetic acid,3-{[4-(3,3-dibenzyl-ureido)-benzenesulfonyl]-[2-(4-methoxy-phenyl)-ethyl]-amino}-N-hydroxy-propionamide,7-(4-methyl-piperazin-1-ylmethyl)-5-phenylsulfanylmethyl-quinolin-8-ol,4-nitro-quinolin-8-ol, 5-butoxymethyl-quinolin-8-ol,[(3-hydroxy-pyridine-2-carbonyl)-amino]-acetic acid,N-((3-hydroxy-6-isopropoxy-quinoline-2-carbonyl)-amino)-acetic acid,[(3-hydroxy-6-trifluoromethoxy-quinoline-2-carbonyl)-amino]-acetic acid,N-((6-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid,((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid methyl ester,N-((7-benzyloxy-1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid, N-((7-butoxy-4-hydroxy-isoquinoline-3-carbonyl)-amino)-aceticacid, 6-cyclohexyl-1-hydroxy-4-methyl-1H-pyridin-2-one,[(6-chloro-3-hydroxy-quinoline-2-carbonyl)-amino]-acetic acid,[(3-methoxy-pyridine-2-carbonyl)-amino]-acetic acid,5-methoxy-4-oxo-1,4-dihydro-[1,1]phenanthroline-3-carboxylic acid,[(1,7-dichloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,{[4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid, and[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid.

In a particular embodiment, a compound for use in the present methods isselected from the group consisting ofN-((1-chloro-4-hydroxy-isoquinoline-3-carbonyl)-amino)-acetic acid,[(1,7-dichloro-4-hydroxy-isoquinoline-3-carbonyl)-amino]-acetic acid,[(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid,{[4-hydroxy-7-(4-methoxy-phenoxy)-isoquinoline-3-carbonyl]-amino}-aceticacid,[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-aceticacid

Pharmaceutical Formulations and Routes of Administration

An effective dose or amount of an agent of the present invention refersto an amount or dose of the agent that results in amelioration ofsymptoms, for example, or an improvement in kidney function, or aprolongation of survival in a subject. An improvement in kidney functioncan be measured by an alteration in any measurable parameter of kidneyfunction, including any of the parameters described herein. An effectiveamount can readily be determined by routine experimentation using avariety of techniques well-known in the art. The effective amount ortherapeutically effective amount is the amount of the agent orpharmaceutical composition that will elicit the desired biological ormedical response that is being sought by the researcher, veterinarian,medical doctor, or other clinician.

The compositions of the present invention can be delivered directly orin pharmaceutical compositions containing carriers, excipients, buffers,etc., as is well known in the art. Pharmaceutically acceptableexcipients are available in the art, and include those listed in variouspharmacopoeias. (See, e.g., USP, JP, EP, and BP, FDA web page(www.fda.gov), Inactive Ingredient Guide 1996, and Handbook ofPharmaceutical Additives, ed. Ash; Synapse Information Resources, Inc.2002.) Various formulations and drug delivery systems are available inthe art. (See, e.g., Gennaro, ed. (2000) Remington's PharmaceuticalSciences, supra; and Hardman, Limbird, and Gilman, eds. (2001) ThePharmacological Basis of Therapeutics.)

The compositions of the present invention can be delivered directly orin pharmaceutical compositions along with suitable carriers orexcipients, as is well known in the art. Present methods of treatmentcan comprise administration of an effective amount of a compound of theinvention to a subject. In a preferred embodiment, the subject is amammalian subject, and in a most preferred embodiment, the subject is ahuman subject. Preferred routes of administration include oral andtransdermal delivery mechanisms.

An effective amount of such agents can readily be determined by routineexperimentation, as can the most effective and convenient route ofadministration and the most appropriate formulation. Variousformulations and drug delivery systems are available and selection of anappropriate formulation is within the level of skill in the art. (See,e.g., Gennaro, ed. (1995) Remington's Pharmaceutical Sciences, supra;and Hardman, Limbird, and Gilman, eds. (2001) The Pharmacological Basisof Therapeutics, supra.)

Suitable routes of administration may, for example, include oral,rectal, transmucosal, nasal, or intestinal administration and parenteraldelivery, including intramuscular, subcutaneous, intramedullaryinjections, as well as intrathecal, direct intraventricular,intravenous, intraperitoneal, intranasal, or intraocular injections. Theagent or composition thereof may be administered in a local rather thana systemic manner. For example, a suitable agent can be delivered viainjection or in a targeted drug delivery system, such as a depot orsustained release formulation.

The pharmaceutical compositions of the present invention may bemanufactured by any of the methods well-known in the art, such as byconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping, or lyophilizing processes. Asnoted above, the compositions of the present invention can include oneor more physiologically acceptable carriers such as excipients andauxiliaries that facilitate processing of active molecules intopreparations for pharmaceutical use.

Proper formulation is dependent upon the route of administration chosen.For injection, for example, the composition may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHanks's solution, Ringer's solution, or physiological saline buffer. Fortransmucosal or nasal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art. For oral administration, the compounds canbe formulated readily by combining the active compounds withpharmaceutically acceptable carriers well known in the art. Suchcarriers enable the compounds of the invention to be formulated astablets, pills, dragees, capsules, liquids, gels, syrups, slurries,suspensions and the like, for oral ingestion by a subject. The compoundsmay also be formulated in rectal compositions such as suppositories orretention enemas, e.g., containing conventional suppository bases suchas cocoa butter or other glycerides.

Pharmaceutical preparations for oral use can be obtained as solidexcipients, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired,disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

Pharmaceutical preparations for oral administration include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added.

In one embodiment, the compounds of the present invention can beadministered transdermally, such as through a skin patch, or topically.In one aspect, the transdermal or topical formulations of the presentinvention can additionally comprise one or multiple penetrationenhancers or other effectors, including agents that enhance migration ofthe delivered compound. Transdermal or topical administration could bepreferred, for example, in situations in which location specificdelivery is desired.

For administration by inhalation, the compounds for use according to thepresent invention are conveniently delivered in the form of an aerosolspray presentation from pressurized packs or a nebuliser, with the useof a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, orany other suitable gas. In the case of a pressurized aerosol, theappropriate dosage unit may be determined by providing a valve todeliver a metered amount. Capsules and cartridges of, for example,gelatin, for use in an inhaler or insufflator may be formulated. Thesetypically contain a powder mix of the compound and a suitable powderbase such as lactose or starch.

Compositions formulated for parenteral administration by injection,e.g., by bolus injection or continuous infusion, can be presented inunit dosage form, e.g., in ampoules or in multi-dose containers, with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Formulations for parenteral administration include aqueoussolutions or other compositions in water-soluble form.

Suspensions of the active compounds may also be prepared as appropriateoily injection suspensions. Suitable lipophilic solvents or vehiclesinclude fatty oils such as sesame oil and synthetic fatty acid esters,such as ethyl oleate or triglycerides, or liposomes. Aqueous injectionsuspensions may contain substances that increase the viscosity of thesuspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, the suspension may also contain suitablestabilizers or agents that increase the solubility of the compounds toallow for the preparation of highly concentrated solutions.Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

As mentioned above, the compositions of the present invention may alsobe formulated as a depot preparation. Such long acting formulations maybe administered by implantation (for example, subcutaneous orintramuscular) or by intramuscular injection. Thus, for example, thepresent compounds may be formulated with suitable polymeric orhydrophobic materials (for example as an emulsion in an acceptable oil)or ion exchange resins, or as sparingly soluble derivatives, forexample, as a sparingly soluble salt.

Suitable carriers for the hydrophobic molecules of the invention arewell-known in the art and include co-solvent systems comprising, forexample, benzyl alcohol, a nonpolar surfactant, a water-miscible organicpolymer, and an aqueous phase. The co-solvent system may be the VPDco-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v ofthe nonpolar surfactant polysorbate 80, and 65% w/v polyethylene glycol300, made up to volume in absolute ethanol. The VPD co-solvent system(VPD:5W) consists of VPD diluted 1:1 with a 5% dextrose in watersolution. This co-solvent system is effective in dissolving hydrophobiccompounds and produces low toxicity upon systemic administration.Naturally, the proportions of a co-solvent system may be variedconsiderably without destroying its solubility and toxicitycharacteristics. Furthermore, the identity of the co-solvent componentsmay be varied. For example, other low-toxicity nonpolar surfactants maybe used instead of polysorbate 80, the fraction size of polyethyleneglycol may be varied, other biocompatible polymers may replacepolyethylene glycol, e.g.,polyvinyl pyrrolidone, and other sugars orpolysaccharides may substitute for dextrose.

Alternatively, other delivery systems for hydrophobic molecules may beemployed. Liposomes and emulsions are well known examples of deliveryvehicles or carriers for hydrophobic drugs. Liposomal delivery systemsare discussed above in the context of gene-delivery systems. Certainorganic solvents such as dimethylsulfoxide also may be employed,although usually at the cost of greater toxicity. Additionally, thecompounds may be delivered using sustained-release systems, such assemi-permeable matrices of solid hydrophobic polymers containing theeffective amount of the composition to be administered. Varioussustained-release materials are established and available to those ofskill in the art. Sustained-release capsules may, depending on theirchemical nature, release the compounds for a few weeks up to over 100days. Depending on the chemical nature and the biological stability ofthe therapeutic reagent, additional strategies for protein stabilizationmay be employed.

For any composition used in the present methods of treatment, atherapeutically effective dose can be estimated initially using avariety of techniques well known in the art. For example, based oninformation obtained from a cell culture assay, a dose can be formulatedin animal models to achieve a circulating concentration range thatincludes the IC₅₀. Similarly, dosage ranges appropriate for humansubjects can be determined, for example, using data obtained from cellculture assays and other animal studies.

A therapeutically effective dose of an agent refers to that amount ofthe agent that results in amelioration of symptoms or a prolongation ofsurvival in a subject. Toxicity and therapeutic efficacy of suchmolecules can be determined by standard pharmaceutical procedures incell cultures or experimental animals, e.g., by determining the LD₅₀(the dose lethal to 50% of the population) and the ED₅₀ (the dosetherapeutically effective in 50% of the population). The dose ratio oftoxic to therapeutic effects is the therapeutic index, which can beexpressed as the ratio LD₅₀, ED₅₀. Agents that exhibit high therapeuticindices are preferred.

Dosages preferably fall within a range of circulating concentrationsthat includes the ED₅₀ with little or no toxicity. Dosages may varywithin this range depending upon the dosage form employed and the routeof administration utilized. The exact formulation, route ofadministration, and dosage should be chosen, according to methods knownin the art, in view of the specifics of a subject's condition.

Dosage amount and interval may be adjusted individually to provideplasma levels of the active moiety that are sufficient to modulate HIFαstabilization and HIF-regulated gene induction, as desired, i.e.,minimal effective concentration (MEC). The MEC will vary for eachcompound but can be estimated from, for example, in vitro data. Dosagesnecessary to achieve the MEC will depend on individual characteristicsof the compound and the route of administration. Agents or compositionsthereof should be administered using a regimen which maintains plasmalevels above the MEC for about 10-90% of the duration of treatment,preferably about 30-90% of the duration of treatment, and mostpreferably between 50-90%. In cases of local administration or selectiveuptake, the effective local concentration of the drug may not be relatedto plasma concentration.

The amount of agent or composition administered will, of course, bedependent on a variety of factors, including the sex, age, and weight ofthe subject being treated, the severity of the affliction, the manner ofadministration, and the judgment of the prescribing physician.

The present compositions may, if desired, be presented in a pack ordispenser device containing one or more unit dosage forms containing theactive ingredient. Such a pack or device may, for example, comprisemetal or plastic foil, such as a blister pack. The pack or dispenserdevice may be accompanied by instructions for administration.Compositions comprising a compound of the invention formulated in acompatible pharmaceutical carrier may also be prepared, placed in anappropriate container, and labeled for treatment of an indicatedcondition.

Methods of Using the Compounds of the Invention

In one aspect, a compound of the invention inhibits one or more2-oxoglutarate dioxygenase enzymes. In one embodiment, the compoundinhibits at least two 2-oxoglutarate dioxygenase family members, e.g.,HIF prolyl hydroxylase and HIF asparagine-hydroxylase (FIH-1), witheither the same specificity or with differential specificity. In anotherembodiment, the compound is specific for one 2-oxoglutarate dioxygenase,e.g., HIF prolyl hydroxylase, and shows little to no specificity forother family members.

The compounds can be administered in combination with various othertherapeutic approaches. In one embodiment, the compound is administeredwith another 2-oxoglutarate dioxygenase inhibitor, wherein the twocompounds have differential specificity for individual 2-oxoglutaratedioxygenase family members. The two compounds may be administered at thesame time as a ratio of one relative to the other. Determination of aratio appropriate to a given course of treatment or a particular subjectis within the level of skill in the art. Alternatively, the twocompounds may be administered consecutively during a treatment timecourse. In a particular embodiment, one compound specifically inhibitsHIF prolyl hydroxylase enzyme activity, and a second compoundspecifically inhibits procollagen prolyl 4-hydroxylase enzyme activity.In another specific embodiment, one compound specifically inhibits HIFprolyl hydroxylase enzyme activity, and a second compound specificallyinhibits HIF asparaginyl-hydroxylase enzyme activity. In anotherembodiment, the compound is administered with another therapeutic agenthaving a different mode of action, e.g., an ACE inhibitor (ACEI),angiotensin-II receptor blocker (ARB), statin, diuretic, digoxin,carnitine, etc.

Compound Screening and Identification

Various assays and screening techniques, including those describedbelow, can be used to identify small molecules that modulate (e.g.,increase or decrease) the level or activity of HIFα. Assays willtypically provide for detectable signals associated with the consumptionof a reaction substrate or production of a reaction product. Detectioncan involve, for example, fluorophores, radioactive isotopes, enzymeconjugates, and other detectable labels well known in the art. Theresults may be qualitative or quantitative. Isolation of the reactionproduct may be facilitated by a label, such as biotin or a histidine tagthat allows purification from other reaction components viaprecipitation or affinity chromatography.

Assays for HIFα hydroxylation may involve measuring hydroxylated prolineor lysine residues in HIFα or a fragment thereof (see, e.g., Palmeriniet al. (1985) J Chromatogr 339:285-292), or measuring formation ofsuccinate from 2-oxoglutarate in the presence of enzyme and HIFα or afragment thereof (see, e.g., Cunliffe et al. (1986) Biochem J240:617-619). Exemplary procedures that measure HIFα hydroxylation aredescribed in Ivan et al. (supra) and Example 8. An exemplary procedurethat measures production of succinate from 2-oxoglutarate is describedby Kaule and Gunzler. (1990; Anal Biochem 184:291-297.) Substratemolecules may include HIFα or a fragment thereof, e.g., HIF(556-575);for example, an exemplary substrate for use in the assay described inExample 8 is [methoxycoumarin]-DLDLEALAPYIPADDDFQL-amide. Enzyme mayinclude, e.g., HIFα prolyl hydroxylase (see, e.g., GenBank Accession No.AAG33965, etc.), obtained from any source. Enzyme may also be present ina crude cell lysate or in a partially purified form. Compounds thatstabilize HIFα or that inhibit hydroxylation of HIFα may be identifiedby measuring and comparing enzyme activity in the absence and presenceof the compound.

Additionally and in combination with the above methods, compounds can beidentified by any of a variety of screening techniques known in the art.Such screening methods may allow for target polypeptides or thecompounds to be free in solution, affixed to a solid support, borne on acell surface, or located within a cell. For example, test compounds maybe arrayed on a surface and analyzed for activity in a manner analogousto array methods currently available in the art. (See, e.g., Shalon etal. (1995) International Publication No. WO 95/35505; Baldeschweiler etal. (1995) International Publication No. WO 95/251116; Brennan et al.(1995) U.S. Pat. No. 5,474,796; and Heller et al. (1997) U.S. Pat. No.5,605,662.)These and other embodiments of the present invention willreadily occur to those of ordinary skill in the art in view of thedisclosure herein, and are specifically contemplated.

EXAMPLES

The invention is understood by reference to the following examples,which are intended to be purely exemplary of the invention. The presentinvention is not limited in scope by the exemplified embodiments, whichare intended as illustrations of single aspects of the invention only.Any methods that are functionally equivalent are within the scope of theinvention. Various modifications of the invention in addition to thosedescribed herein will become apparent to those skilled in the art fromthe foregoing description and accompanying figures. Such modificationsfall within the scope of the appended claims.

Example 1 Increased GFR and Decreased Serum Creatinine Levels in HumanSubjects

Patients with chronic kidney disease (CKD), and with anemia associatedwith CKD, not yet receiving dialysis and not receiving recombinant EPOtherapy, were randomized to either a treatment group or a placebo group.The treatment group was administered compound B (6 mg/kg) orally threetimes a week (Monday, Wednesday, and Friday) for four weeks (i.e., day 0to day 28). Compound B has been previously shown to both stabilize HIFαand to inhibit HIF hydroxylase activity. Both groups were observed foran additional two weeks (i.e., to day 42). Glomerular filtration ratewas estimated as standard in the art from serum creatinine measurementsusing the Cockcroft-Gault formula. (See Cockcroft and Gault (1976)Nephron 16:31-41.)

Mean estimated GFR for all patients on day 0 was 20.0 ml/min/1.73 m2(range=12.4 to 31.7 ml/min/1.73 m²), indicating severe renal impairmentand reduced renal function associated with chronic kidney disease. Asshown in FIG. 1A, administration of a compound of the present inventionincreased eGFR in patients (e.g., patient 04-004 and patient 04-009). Nochange in eGFR was observed in the placebo group (FIG. 1B, patient04-003 and patient 04-008). Table 1 below shows serum creatinine levels(μmol/l) in patients treated with compound B or with placebo. The valuesfor serum creatinine levels (μmol/l) shown in Table 1 can be expressedas mg/dl. For example, a serum creatinine level of 231 mmol/l (patient04-004, day 42) can be expressed as 2.625 mg/dl. As shown in Table 1below, patients treated with compound B (e.g., patient 04-004 andpatient 04-009) showed decreased serum creatinine levels.

TABLE 1 Day Day Day Day Day 0 Day 7 Day 14 21 28 35 42 Patient 04-001647 637 706 761 711 703 716 Patient 04-002 316 373 295 314 331 340 411Patient 04-003 404 413 430 428 (placebo) Patient 04-004 376 305 284 257275 231 Patient 04-008 397 365 432 414 425 393 416 (placebo) Patient0-009 424 432 360 489 455 365 Patient 04-106 417 570 449 441 425

These results demonstrated that methods and compounds of the presentinvention increased GFR, and decreased serum creatinine, in patientshaving severe renal impairment and chronic kidney disease. Therefore,the present methods and compounds are effective in improving kidneyfunction in human subjects.

Example 2 Improved Renal Function in a Rat Model of Impaired KidneyFunction

To examine the effects of compounds and methods of the present inventionon kidney function, the following studies were performed. For thesestudies, male Sprague-Dawley rats (Winkelmann, Borchen, Germany) wereused at weights of 180-230 grams. Animals were fed a standard diet andhad free access to water. In these studies, animals were administered(by intraperitoneal injection) a dose of 25 mg/kg compound R, dissolvedin 100 μl DMSO and 900 μl NaCl (0.9%), 6 hours prior to initiation ofthe ischemic insult, as described below. In some experiments, animalswere exposed to 0.1% carbon monoxide (CO) as previously described. (SeeRosenberger et al (2002) J Am Soc Nephrol 13:1721-1732.)

Kidney dysfunction (impaired kidney function) was induced by renalischemia-reperfusion injury (IRI) as follows. Animals were anaesthetizedwith ketamine (100 mg/kg) and pentobarbital sodium (50 mg/kg, Nembutal®,Abbott, Wiesbaden, Germany). A constant body temperature of 37° C. wasmaintained using a heated table with temperature feedback by a rectalprobe (Heating Conroller Type 861, Hugo Sachs Elektronik-HarvardApparatus GmbH, March, Germany). A blood sample of 300 μl was taken viatail vein puncture (time point 0 hr). The abdomen was shaved and amidline incision was made. After removal of the right kidney, the leftrenal artery was clamped with an arterial clamp (B-2, S&T, Neuhausen,Switzerland) to induce ischemia in the kidney, which was verified by thechange of the renal color. After an ischemic period of 40 minutes, thearterial clamp was removed and reperfusion of the kidney was initiated.The abdomen was closed and animals were kept on the heating table untilawakening after anaesthesia. Twenty-four hours later, the animals werebriefly anaesthetized with isoflourane (Forene®, Abbott, Wiesbaden,Germany) and a second blood sample (24 hr) of 300 μl was drawn. After 72hr the animals were anaesthetised and a third blood sample (72 hr) wasdrawn.

For these experiments, animals were divided into five treatment groups(n=10 for each group) as follows: Group A (Sham), sham operated animalswithout impaired kidney function; Group B (CO), animals treated with0.1% carbon monoxide prior to induction of impaired kidney function;Group C (UnT), untreated animals with IRI; Group D (Compound R), animalstreated with compound R 6 hours prior to induction of impaired kidneyfunction; and Group E (Veh), vehicle-treated animals injected with 100μl DMSO+900 μl NaCl (0.9%) 6 hours prior to induction of impaired kidneyfunction.

Serum Urea

As shown in Table 2 below, both untreated control animals (UnT) andvehicle-treated control animals (Veh) had increased serum urea levels 24hours and 72 hours following the induction of impaired renal functioncompared to that seen in sham control animals. This data indicated thatrenal injury resulted in reduced kidney function, as evidenced byincreased serum urea levels. Animals exposed to 0.1% CO for 10 hours ortreated with compound B 6 hours prior to induction of impaired renalfunction had reduced levels of serum urea compared to untreated (UnT) orvehicle-treated (Veh) control animals. Specifically, 24 hours and 72hours after induction of impaired renal function, serum urea levels weresignificantly lower in animals administered compound B compared tountreated control animals. (See Table 2; data represents mean serum urealevels±standard deviation; #=p<0.05 vs. sham; *=p<0.05 vs. UnT or Veh;n=10.) This data indicated that administration of compound B improvedrenal function compared to untreated and vehicle-treated controlanimals.

TABLE 2 Serum urea Serum urea Serum urea [mg/dl] [mg/dl] [mg/dl] 0 hr 24hr 72 hr Sham 37 +/− 7  46 +/− 6  46 +/− 5 (Group A) UnT 39 +/− 10 257+/− 40 # 202 +/− 70 # (Group B) CO 52 +/− 8 158 ( 78 # * 120 ( 78 # *(Group C) Veh 35 +/− 10 319 +/− 135 # 265 +/− 113 # (Group D) Cmpd R 41+/− 10 179 +/− 61 # *  91 +/− 36 # * (Group E)

Serum Creatinine

As shown in FIGS. 2A and 2B, both untreated control animals (UnT) andvehicle-treated control animals (Veh) had increased serum creatininelevels 24 hours and 72 hours following the renal ischemic injurycompared to that seen in sham control animals. This data indicated thatinduction of impaired renal function resulted in reduced kidneyfunction, as evidenced by increased serum creatinine levels. Animalsexposed to 0.1% CO for 10 hours prior to induction of impaired renalfunction or treated with compound R 6 hours prior to induction ofimpaired renal function had reduced levels of serum creatinine comparedto untreated (UnT) or vehicle-treated (Veh) control animals.Specifically, 24 hours and 72 hours after onset of kidney dysfunction,serum creatinine levels were significantly lower in animals administeredcompound R compared to untreated control animals. (See FIGS. 2A and 2B;data represents mean serum creatinine levels±standard deviation;#=p<0.05 vs. sham; *=p<0.05 vs. UnT or Veh; n=10.) This data showed thatadministration of compound R improved renal function compared tountreated and vehicle-treated control animals. These results indicatedthat methods and compounds of the present invention were effective atimproving kidney function.

Example 3 Improved Renal Function in a Rat Model of Impaired KidneyFunction

To examine the effects of compounds and methods of the present inventionon kidney function, the following studies were performed. For thesestudies, Sprague Dawley rats (Charles River Labs) were used. Animalswere housed in an animal facility according to IACUC protocols with freeaccess to water and food, and all experiments conducted according to theNational Institutes of Health guidelines for animal experimentation.

Renal ischemia-reperfusion injury (IRI) and renal dysfunction wereinduced in male Sprague-Dawley rats (280-300 g) as previously described.(See Nemoto et al (2001) Kidney Int 59:246-251.) Briefly, rats wereanesthetized under isoflurane and a midline abdominal incision was madeunder sterile conditions followed by bluntly dissecting the renalpedicles. A vascular clip was placed on the right renal pedicle whilethe left kidney underwent simultaneous nephrectomy. After 45 minutes ofocclusion (ischemia), the clip was released and reperfusion was observedby changing color of the kidney. Animal body temperature was maintainedconstant, and warm 0.9% saline (0.5% of body weight) was administereddirectly into abdomen before the incision was completely sutured. Sixhours before ischemia-reperfusion injury was performed, rats wereadministered either equal volumes of 0.5% CMC or 135 mg/kg compound S insuspension by oral gavage in a volume of 6 ml/kg.

Blood was collected on days 3, 7 and 10 after IRI for measurement ofblood urea nitrogen (BUN) and serum creatinine levels. After warmingunder a heating lamp, rats were anesthetized with isoflurane and bloodcollected from the tail vein. For serum analysis, −0.7 ml of blood wastransferred to a Microtainer Serum Separator tube followed by incubationat room temperature for −30 minutes, with centrifugation at 10,000 rpmin a Eppendorf 5415C Micro Centrifuge (Hayward, Calif.) for 10 min at 4°C.

BUN and serum creatinine concentrations were measured using an automatedanalyzer (Roche/Hitachi 911). BUN concentration was determined by anenzymatic procedure according to the method of Talke and Shubert. (SeeTalke and Schubert (1965) Klin Wochenschr 43:174-175.) Serum creatinineconcentration was determined by the Jaffe reaction as modified byBartels. (See Bartels and Bohmer (1973) Med Lab (Stuttg.) 26:209-215.)For BUN measurements, the inter-assay coefficient of variance (CV) andthe intra-assay CV were determined to be <3.6% and <2.1%, respectively.For serum creatinine measurements, the inter-assay CV and theintra-assay CV were determined to be <2.1% and <1.7%, respectively.

As shown in FIG. 3, animals administered vehicle control prior to renalIRI had statistically significant increases in BUN and serum creatininelevels on day 3 compared to sham operated animals without IRI (BUN, shamvs IRI, p<0.05; serum creatinine, sham vs. IR, p<0.05). Elevated levelsof both BUN and serum creatinine peaked at day 3 following induction ofimpaired renal function and gradually diminished over time, but remainedabove that of sham control animals at day 7 and day 10 post-IRI. Incontrast to animals administered vehicle and undergoing IRI, animalsadministered compound S 6 hours prior to induction of renal dysfunctionshowed a statistically significant reduction in peak BUN and serumcreatinine levels at day 3, a significant reduction in BUN at day 7 andday 10, and significant reduction in serum creatinine levels at day 7(p<0.05 for each comparison). Specifically, these results showed that asingle dose of compound S administered six hours prior to induction ofimpaired renal function improved kidney function as evidenced bysustained reductions in both serum creatinine levels (day 3 post MRI,2.37±0.59 mg/dl vs. IRI+Cmpd C, 0.73±0.21 mg/dl, p<0.004) and blood ureanitrogen (BUN) (day 3 post IRI, 112.8±23 mg/dl vs. IRI+Cmpd C, 43±12mg/dl, p<0.004). These results showed that compound S administered toanimals 6 hours prior to induction of impaired renal function waseffective at improving kidney function. These results indicated thatmethods and compounds of the present invention were effective atimproving kidney function.

Example 4 Improved Kidney Function in a Mouse Model of Impaired KidneyFunction

To examine the effects of compounds and methods of the present inventionon kidney function, the following studies were performed. In this study,the effects of oral administration of compound D on renal function in amouse model of renal IRI were examined. Male C57BL/6 mice (Charles RiverLaboratories, Hollister, Calif.) of approximately 22-25 grams were usedin these studies.

Renal ischemia-reperfusion injury (IRI) and renal dysfunction wereinduced in mice using techniques previously described. (See Patel et al(2004) Kidney Int 66(3):983-989 and Lee et al (2004) J Am Soc Nephrol15:102-111.) Briefly, mice were anesthetized by intraperitoneal (i.p.)injection of chloral hydrate at 300-400 mg/kg (8-10 mL/kg of 4%) or toeffect. Kidneys were exposed through an abdominal section, and the rightkidney was removed after its vascular pedicle and ureter were ligated.The vascular pedicle of the left kidney was clamped by a micro-aneurysmclip for 40 minutes after right nephrectomy. After the renal clamp wasremoved, the kidney was observed for reflow (reperfusion) after which0.6 mL saline at 37° C. was injected into the abdomen and the incisionwas sutured. Mice were kept on a heating blanket and allowed to recoverfrom anesthesia. All experimental techniques were in accordance toNational Institutes of Health guidelines for animal experimentation.

For these studies, animals were divided into 5 treatment groups asfollows. Group A, oral vehicle administered at 10 mL/kg withoutinduction of impaired renal function; Group B, oral vehicle administeredat 10 mL/kg 6 hours prior to induction of impaired renal function; GroupC, oral administration of compound T (15 mg/kg) 6 hours prior toinduction of impaired renal function; Group D, oral administration ofcompound T (30 mg/kg) 6 hours prior to induction of impaired renalfunction; and Group E, oral administration of compound T (60 mg/kg) 6hours prior to induction of impaired renal function. All treatmentgroups administered compound T were treated with a single oraladministration of compound T 6 hours prior to induction of renaldysfunction.

Twenty-four hours after induction of impaired renal function, blood wascollected from the animals from the abdominal vein (animals underisoflurane anesthesia) into a heparinized tube for plasma preparation.The plasma tubes were centrifuged for 10 min at 4° C. and the plasma wastransferred to a 1.5 ml tube. The plasma was transferred on wet ice toQuality Clinical Labs, Inc. (Mountain View, Calif.) for analysis of BUNlevels and serum creatinine levels.

As shown in FIG. 4, animals administered vehicle control prior toinduction of impaired renal function (Group B) had elevated BUN levelsone day after induction of impaired renal function compared to that ofsham control animals (Group A). Animals administered compound T prior toinduction of impaired renal function showed significantly reduced BUNlevels compared to vehicle-treated animals with renal dysfunction.Specifically, administration of various concentrations of compound T(Group C, 15 mg/kg; Group D, 30 mg/kg; or Group E, 60 mg/kg) improvedrenal function as demonstrated by reducing the elevated BUN levels 24hours after induction of impaired renal function. (Values in FIG. 4 arepresented as mean values for BUN levels±SEM.) (* significantly higherthan Group A; # significantly lower than Group B.)

As shown in FIG. 5, animals administered vehicle control prior toinduction of impaired renal function (Group B) had elevated serumcreatinine levels one day after induction of impaired renal functioncompared to that of sham control animals (Group A). Animals administeredcompound T prior to induction of impaired renal function showedsignificantly reduced serum creatinine levels compared tovehicle-treated animals with renal dysfunction. Specifically,administration of various concentrations of compound T (Group C, 15mg/kg; Group D, 30 mg/kg; or Group E, 60 mg/kg) improved renal functionas demonstrated by reducing the elevated serum creatinine levels 24hours after induction of impaired renal function. (Values in FIG. 5 arepresented as mean values for serum creatinine levels±SEM.) (*significantly higher than Group A; # significantly lower than Group B.)

These results demonstrated that methods and compounds of the presentinvention were effective at improving renal function.

Example 5 Improved Kidney Function in an Animal Model of Impaired KidneyFunction

To examine the effects of compounds and methods of the present inventionon kidney function, the following studies were performed. In this study,the effects of intravenous (i.v.) administration of compound T on renalfunction in a rat model of renal IRI were examined. Additionally, theeffects of compound T administered at various times prior to andfollowing induction of impaired renal function were also examined. Forthese studies, male Sprague Dawley rats (Charles River Labs) ofapproximately 280-300 grams were used. Animals were housed in an animalfacility according to IACUC protocols with free access to water andfood, and all experiments conducted according to the National Institutesof Health guidelines for animal experimentation.

Renal ischemia-reperfusion injury (IRI) and renal dysfunction wereinduced in rats using techniques previously described. (See Nemoto et al(2001) Kidney Int 59:246-251.) Briefly, rats were anesthetized underisoflurane and a midline abdominal incision was made under sterileconditions followed by bluntly dissecting the renal pedicles. Amicrovascular clamp was placed on the left renal pedicle for 45 minuteswhile the night kidney underwent simultaneous nephrectomy. After eachocclusion, the clip was released at 45 minutes, and reperfusion wasconfirmed by observing a change in color of the kidney. Temperature wasmaintained constant, and warm (˜37° C.) saline (1.0 mL) was administereddirectly into the abdomen. After the renal clamp was removed, theincision was sutured and the animal was allowed to recover and had freeaccess to food and water. All experimental techniques were in accordanceto National Institutes of Health guidelines for animal experimentation.

For these studies, animals were divided into 5 treatment groups and weretreated with a single intravenous administration of vehicle or compoundT as follows. Group A, i.v. administration of vehicle (6 mL/kg) 4 hourspre-sham operation without kidney dysfunction; Group B, i.v.administration of vehicle 4 hours prior to induction of impaired kidneyfunction; Group C, i.v. administration of Cmpd T (20 mg/kg) 4 hoursprior to induction of impaired kidney function; Group D, i.v.administration of Cmpd T (20 mg/kg) 45 minutes following induction ofimpaired kidney function; and Group E, i.v. administration of Cmpd T (20mg/kg) 2 hours following induction of impaired kidney function.

Blood was collected on day 3, day 7, and day 10 for analysis of BUNlevels and serum creatinine levels. After warm up under a heating lamp,rats were placed under isoflurane anesthesia and blood was collectedfrom the tail vein at day 3 and day 7 and from the abdominal main veinat day 10. For serum, about 0.6 ml of blood was transferred to aMicrotainer Serum Separator tube (Becton-Dickinson#365960). After beingleft at room temperature for about 30 minutes, the tubes werecentrifuged at 4° C. and the serum (upper layer) was transferred to a1.5 ml tube. The serum was transferred on wet ice to Quality ClinicalLabs, Inc. (Mountain View, Calif.) for clinical chemistry analysiswithin 12 hours.

As shown in FIG. 6 (day 3), FIG. 7 (day 7), and FIG. 8 (day 10), animalsadministered vehicle control prior to induction of impaired renalfunction (Group B) had elevated BUN levels compared to that of shamcontrol animals (Group A). Values in FIG. 6, FIG. 7, and FIG. 8 arepresented as mean values for BUN levels±SEM. At day 3 (FIG. 6), day 7(FIG. 7), and day 10 (FIG. 8), animals administered compound T 4 hoursprior to IRI (Group C), 45 minutes following impairment of renalfunction (Group D), or 2 hours following impairment of renal function(Group E) had decreased BUN levels compared to vehicle-treated controlanimals with kidney dysfunction (Group B). These results showed thatadministration of compound T before or after induction of renaldysfunction improved renal dysfunction as demonstrated by a reduction inthe elevated BUN levels. These results indicated that methods andcompounds of the present invention are effective at reducing BUN levelsand improving kidney function. (FIG. 6, * significantly higher thanGroup A; FIG. 7, * significantly higher than Group A (p<0.001); FIG.8, * significantly higher than Group A; FIG. 8, # significantly lowerthan Group B.)

As shown in FIG. 9 (day 3), FIG. 1 (day 7), and FIG. 11 (day 10),animals administered vehicle control prior to induction of impairedrenal function (Group B) had elevated serum creatinine levels comparedto that of sham control animals (Group A). Values in FIG. 9, FIG. 10,and FIG. 11 are presented as mean values for serum creatininelevels±SEM. At day 3 (FIG. 9), day 7 (FIG. 10), and day 10 (FIG. 11),animals administered compound T 4 hours prior to induction of impairedrenal function (Group C), 45 minutes following induction of impairedrenal function (Group D), or 2 hours following induction of impairedrenal function (Group E) had decreased serum creatinine levels comparedto vehicle-treated control animals with kidney dysfunction (Group B).These results showed that administration of compound T before or afterinduction of renal dysfunction improved renal dysfunction asdemonstrated by a reduction in the elevated serum creatinine levels.These results indicated that methods and compounds of the presentinvention are effective at reducing serum creatinine levels andimproving kidney function. (FIG. 9, * significantly higher than Group A;FIG. 9, # significantly lower than Group B; FIG. 10, * significantlyhigher than Group A; FIG. 10, # significantly lower than Group B; FIG.11, * significantly higher than Group A; FIG. 1, # significantly lowerthan Group B.)

Example 6 Improved Kidney Function in an Animal Model ofChemotherapy-Induced Kidney Dysfunction

To examine the effects of compounds and methods of the present inventionon kidney function following chemotherapy-induced renal injury, thefollowing studies were performed. For the studies described below, maleSprague Dawley rats (Charles River Labs) of approximately 280-300 gramswere used. Animals were housed in an animal facility according to IACUCprotocols with free access to water and food, and all experimentsconducted according to the National Institutes of Health guidelines foranimal experimentation.

Cisplatin (CP) is an effective chemotherapeutic agent used in thetreatment of a variety of solid tumors. Administration of CP is oftenassociated with kidney injury and nephrotoxicity, resulting in impairedkidney function. For these experiments, CP-induced kidney dysfunctionwas performed as previously described (See Bagnis et al (2001) NephrolKial Transplant 16:932-938), with the following modifications. A singledose of CP (Cisplatin Injection, obtained from Bedford Laboratories,Bedford, Ohio) at 1.0 mg/mL was administered i.v. on day 0. Animals wereadministered compound B or compound U by oral gavage on day 0, day 2,day 4, day 7, day 9, and day 11. The initial dose of compound B orcompound U was given ˜2 h before the i.v. injection of CP.

For these studies, animals were divided into 8 treatment groups follows.Group A, i.v. administration of saline (5 mL/kg) and oral gavageadministration of vehicle (5 mL/kg) 3×/wk; Group B, i.v. administrationof saline (5 mL/kg) and oral gavage administration of Cmpd B (60 mg/kg)3×/wk; Group C, i.v. administration of saline (5 mL/kg) and oral gavageadministration of Cmpd U (40 mg/kg) 3×/wk; Group D, i.v. administrationof CP (5 mg/kg) and oral gavage administration of vehicle (5 mL/kg)3×/wk; Group E, i.v. administration of CP (5 mg/kg) and oral gavageadministration of Cmpd B (30 mg/kg) 3×/wk;Group F, i.v. administrationof CP (5 mg/kg) and oral gavage administration of Cmpd B (60 mg/kg)3×/wk;Group G, i.v. administration of CP (5 mg/kg) and oral gavageadministration of Cmpd U (20 mg/kg) 3×/wk; and Group H, i.v.administration of CP (5 mg/kg) and oral gavage administration of Cmpd U(40 mg/kg) 3×/wk.

On day 4, day 7, and day 14, blood was obtained from the animals foranalysis of BUN levels and serum creatinine levels. Blood samples wereobtained as follows. After being warmed under a heating lamp, animalswere placed under isoflurane anesthesia and blood was collected from thetail vein. About 0.5 ml of blood was transferred to a Microtainer SerumSeparator tube (Becton-Dickinson #365960). After being left at roomtemperature for about 30 min, the tubes were centrifuged at 4° C. andthe serum (upper layer) was transferred to a 1.5 ml tube. The serum wastransferred on wet ice to Quality Clinical Labs, Inc. (Mountain View,Calif.) for clinical chemistry analysis within 12 hours.

As shown in FIG. 12 (day 4), FIG. 13, (day 7), and FIG. 14 (day 14),animals administered CP and vehicle control (Group D) had elevated BUNlevels compared to that of control animals (Group A). Values in FIG. 12,FIG. 13, and FIG. 14 are presented as mean values for BUN levels ±SEM.At day 4 (FIG. 12), day 7 (FIG. 13), and day 14 (FIG. 14), animalsinjected with CP and administered compound B (Group E and Group F) orcompound U (Group G and Group H) had decreased BUN levels compared tovehicle-treated animals injected with CP (Group D). These results showedthat administration of compound B or compound U improved renaldysfunction following CP administration, as demonstrated by a reductionin the CP-induced elevation of BUN levels. These results indicated thatmethods and compounds of the present invention are effective at reducingBUN levels and improving kidney function.

As shown in FIG. 15 (day 4), FIG. 16, (day 7), and FIG. 17 (day 14),animals administered CP and vehicle control (Group D) had elevated serumcreatinine levels compared to that of control animals (Group A). Valuesin FIG. 15, FIG. 16, and FIG. 17 are presented as mean values for serumcreatinine levels ±SEM. At day 4 (FIG. 15), day 7 (FIG. 16), and day 14(FIG. 17), animals injected with CP and administered compound B (Group Eand Group F) or compound U (Group G and Group H) had decreased serumcreatinine levels compared to vehicle-treated animals injected with CP(Group D). These results showed that administration of compound B orcompound U improved renal dysfunction following CP administration, asdemonstrated by a reduction in the CP-induced elevation of serumcreatinine levels. These results indicated that methods and compounds ofthe present invention are effective at reducing serum creatinine levelsand improving kidney function.

Example 7 Renal Ischemia-Reperfusion Injury

The model of ischemic acute renal failure was described in Nemoto et al.(2001, Kidney Int 59:246-251.) Briefly, male Sprague-Dawley rats(200-250 g) were treated with either 0.5% carboxymethyl cellulose (CMC;Sigma-Aldrich) or 1.5% compound B suspended in CMC by oral gavage in avolume of 4 ml/kg/day. Rats were pretreated daily for 4 consecutive days(days −3 to 0). A few hours after the fourth and last oral dose on day0, renal ischemia-reperfusion injury (IRI) was performed.

Animals were divided into four groups: (1) Vehicle pretreatment and shamsurgery; (2) compound B pretreatment and sham surgery; (3) vehiclepretreatment and EM surgery; and (4) compound B pretreatment and IRIsurgery. Animals were anesthetized under isoflurane, an incision wasmade in the abdominal midline, and the renal pedicles were bluntlydissected. A vascular clip was placed on the right renal pedicle for 45minutes while the left kidney underwent simultaneous nephrectomy. Aftereach occlusion, the clip was released at 45 minutes, and reperfusion wasobserved by the changing color of the kidney. Temperature was maintainedconstant, and warm saline (0.5% of body weight) containing Buprenexanalgesic was administered directly into abdomen before the incision wascompletely sutured.

The animal body weight and mortality were monitored. Blood samples wereobtained from the tail vein, and serum chemistry and CBC were measuredby IDEXX veterinary service (West Sacramento Calif.). Data are presentedas mean ±SE with number of animals in parenthesis. The data werecompared within the four groups at each time point using one-wayanalysis of variance (ANOVA, SIGMASTAT) and Student-Newman-Keuls method.A value of P<0.05 was considered significant.

As can be seen in FIG. 18, treatment with the compound prevented earlymortality associated with ischemic-reperfusion injury. Further, serumblood urea nitrogen (BUN), a gauge of renal function, was significantlyelevated by renal IRI at both 3 and 7 days, whereas treatment withcompound produced significantly less IRI-induced increase in BUN. (FIG.19A.) Additionally, serum cholesterol was significantly elevated byrenal IRI at days 3, 7 and 14, whereas treatment with compoundcompletely blocked IRI-induced increase in serum cholesterol. (FIG.19B.) Athough the reasons are still under investigation, elevated kidneycholesterol is a natural reflection of renal ischemic-reperfusioninjury. (Zager et al. (2001) Am J Pathol 159:743-752; Appel (1991)Kidney Int 39:169-183; and Abdel-Gayoum et al. (1999) Hum Exp Toxicol18:454459.)

Example 8 Screening Assay

Compounds that inhibit HIF-specific prolyl hydroxylase activity andthereby stabilize HIFα can be identified and characterized using thefollowing assay. A 50 μt aliquot of a reaction mix containing 4 mg/mlBSA, 0.1 M Tris HCl (pH 7.2), 2 mM ascorbate, 80 μM ferrous sulfate, 0.2mM 2-oxoglutarate, 600 units/ml catalase, with or without 100 μM HIFαpeptide is mixed with 50 μl HeLa cell extract or purified HIF prolylhydroxylase and incubated 1.5 hours at 37° C. Following incubation, 50μl of streptavidin beads are added and the mixture is incubated for 1hour with agitation at 4° C. The mixture is transferred to tubes andcentrifuged at low speed to pellet the beads. The beads are washed threetimes with 0.5 to 1 ml 20 mM Tris HCl (pH 7.2). The peptide is theneluted from the beads with 5 μl 2 mM biotin in 20 mM Tris HCl (pH 7.2)for 1 hour. The tubes are centrifuged to pellet the resin and 40-50 μlof supernatant is removed and an equal volume of acetonitrile is added.Alternatively, the peptide is attached to methoxycoumarin, a pHinsensitive fluorophore. The fluorophore may provide sensitivity andspecificity to enhance detection in assays run with crude cell lysate.An exemplary HIF peptide for use in the screening assay may comprise[methoxycoumarin]-DLDLEALAPYIPADDDFQL-amide. The non-hydroxylated andhydroxylated peptides are then separated by reverse-phase HPLC on a C18column with UV detection at 214 nm.

Various modifications of the invention, in addition to those shown anddescribed herein, will become apparent to those skilled in the art fromthe foregoing description. Such modifications are intended to fallwithin the scope of the appended claims.

All references cited herein are hereby incorporated by reference hereinin their entirety.

1. A method for improving kidney function in a subject having impairedkidney function, the method comprising administering to the subject aneffective amount of an agent that inhibits hypoxia inducible factor(HIF) hydroxylase activity.
 2. The method of claim 1, wherein thesubject is a mammalian subject.
 3. The method of claim 1, wherein thesubject is a human subject.
 4. A method for increasing glomerularfiltration rate (GFR) in a subject having a decreased GFR, the methodcomprising administering to the subject an effective amount of an agentthat inhibits HIF hydroxylase activity.
 5. The method of claim 4,wherein the decreased GFR is selected from the group consisting of:below about 116 ml/min/1.73 m²; below about 107 ml/min/1.73 m²; belowabout 99 ml/min/1.73 m²; below about 93 ml/min/1.73 m²; below about 85ml/min/1.73 m²; and below about 75 ml/min/1.73 m².
 6. The method ofclaim 4, wherein the increasing GFR comprises increasing GFR to a levelselected from the group consisting of: above about 15 ml/min/1.73 m²;above about 30 ml/min/1.73 m²; above about 60 ml/min/1.73 m²; and aboveabout 90 ml/min/1.73m².
 7. A method for decreasing serum creatinine in asubject having or at risk for having increased serum creatinine, themethod comprising administering to the subject an effective amount of anagent that inhibits HIF hydroxylase activity.
 8. The method of claim 7,wherein the increased serum creatinine is above a serum creatinine levelselected from the group consisting of: 0.6, 0.8, 1.0, and 1.4 mg/dl. 9.The method of claim 8, wherein the decreasing serum creatinine comprisesdecreasing serum creatinine to below a serum creatinine level selectedfrom the group consisting of: 0.6, 0.8, 1.0, and 1.4 mg/dl.
 10. A methodfor decreasing blood urea nitrogen (BUN) in a subject having or at riskfor having increased BUN levels, the method comprising administering tothe subject an effective amount of an agent that inhibits HIFhydroxylase activity.
 11. The method of claim 10, wherein the increasedBUN is greater than a BUN selected from the group consisting of: 10mg/dl, 20 mg/dl, 30 mg/dl, 40 mg/dl, 50 mg/dl, 60 mg/dl, 70 mg/dl, and80 mg/dl.
 12. The method of claim 10, wherein the decreasing BUNcomprises decreasing BUN to a level at or below about 60 mg/dl.
 13. Amethod for decreasing cholesterol in a subject having or at risk forhaving elevated cholesterol, the method comprising administering to thesubject an effective amount of an agent that inhibits HIF hydroxylaseactivity.
 14. The method of claim 13, wherein the reducing cholesterolcomprises reducing blood cholesterol levels to a level selected from thegroup consisting of: below 200 mg/dL, below 180 mg/dL, below 160 mg/dL,and below 150 mg/dL.
 15. The method of claim 13, wherein the elevatedcholesterol is selected from the group consisting of a blood cholesterollevel above 200 mg/dL above 220 mg/dL, and above 240 mg/dL.
 16. Themethod of claim 1, wherein the subject has or is at risk for having anacute or chronic kidney disease.
 17. The method of claim 1, wherein theacute or chronic kidney disease is acute kidney failure or chronickidney failure.
 18. The method of claim 1, wherein the subject hasdiabetes.
 19. The method of claim 1, wherein the subject hashypertension.
 20. A pharmaceutical composition for improving kidneyfunction, the composition comprising an effective amount of an agentthat inhibits HIF hydroxylase activity.